These findings demonstrate that the +37 kb Cebpa enhancer is central to regulation of Cebpa transcription and granulopoiesis in vivo

Runx1, C/EBPα, Pu.one, Erg, Fli-1, GATA2, Scl, Meis1, and Gfi-1b bind chromatin in the region of this enhancer in hematopoietic cells as determined by ChIP-Seq, Runx1, C/EBPα, Pu.1, Fli-1, Erg, Ets1, c-Myb, GATA2, and Scl bind conserved enhancer cis elementsPF-4708671 distributor in gel shift assays, and mutation of the Runx1, C/EBP, Ets, Myb, GATA, or E-box web-sites every single reduce enhancer action in 32Dcl3 myeloid cells in reporter assays. Mutation of its 7 Ets sites led to the biggest reduction in enhancer activity, and CRISPR/Cas9-mediated substitution of the endogenous enhancer alleles with a variant harboring place mutations in these Ets websites led to 20-fold reduced Cebpa mRNA expression in 32Dcl3 myeloid cells.To ascertain whether or not the +37 kb Cebpa enhancer is also crucial for regulating Cebpa expression in vivo, we have now created and characterized mice in which loxP internet sites flank the enhancer, selected as Enh mice. Germline deletion employing CMV-Cre discovered marked reduction of Cebpa expression in marrow but not in other tissues, which include liver, adipose, and lung, that usually convey C/EBPα. As germline deletion or use of Vav-Cre to induce hematopoietic-distinct deletion led to major early post-natal lethality, we concentrated on examination of grownup EnhMx1-Cre mice subjected to pIpC injections to induce enhancer deletion, followed by restoration for 4 months to reestablish homeostasis and to keep away from transient pIpC effects. In this design, Cebpa mRNA was decreased 14-fold in GMP or CMP and 30-fold in the LSK marrow population associated with a 3-fold reduction in GMP, LSK enlargement, LSK/SLAM cell depletion, and impaired granulopoiesis relative to monopoiesis. Erythroid progenitor and platelet enlargement and decreased figures of B lymphoid colony forming units was also observed, with preservation of useful LT-HSC. These findings reveal that the +37 kb Cebpa enhancer is central to regulation of Cebpa transcription and granulopoiesis in vivo.IndomethacinFor mobile cycle evaluation utilizing the BrdU Stream kit , mice had been provided a one intraperitoneal injection of 5-bromodeoxyuridine . 3 hrs later, bone marrow was isolated from femurs, tibias, iliac crest and backbone. Right after pink mobile lysis, the cells have been lineage depleted as above. Lineage unfavorable cells were stained with Live/Useless Fixable Aqua and for LSK, CMP, GMP, and MEP as over. The fixation, DNase remedy, and staining with FITC anti-BrdU and 7AAD had been for each the BrdU Circulation kit protocol. For quiescence examination, lineage depleted marrow was stained with Are living/Useless Fixable Aqua and floor makers for LSK, CMP, GMP, and MEP, preset, handled with DNase, and stained with FITC-anti-Ki67 and 7AAD.

1,069 thoughts on “These findings demonstrate that the +37 kb Cebpa enhancer is central to regulation of Cebpa transcription and granulopoiesis in vivo

  1. Pingback: party tents
  2. Pingback: euro iptv
  3. Pingback: jobs at home
  4. Pingback: herbals
  5. Pingback: Miami OHIO
  6. Pingback: check that
  7. Pingback: kosher sex toy
  8. Pingback: kona coffee
  9. Pingback: why not try here
  10. Pingback: make your own app
  11. Pingback: ground coffee
  12. Pingback: weight vest
  13. Pingback: how to make a app
  14. Pingback: Miniclip Oyunları
  15. Pingback: build an app
  16. Pingback: jasa anti rayap
  17. Pingback: back links
  18. Pingback: look what i found
  19. Pingback: my site
  20. Pingback: check it out
  21. Pingback: new movies
  22. Pingback: HD 배경 화면
  23. Pingback: acheter like
  24. Pingback: gem residences psf
  25. Pingback: how to earn
  26. Pingback: see it here
  27. Pingback: gourmet coffee
  28. Pingback: kona coffee
  29. Pingback: kona coffee
  30. Pingback: coffee beans
  31. Pingback: bantningspiller
  32. Pingback: container homes
  33. Pingback: gps fleet tracking
  34. Pingback: kona coffee
  35. Pingback: kona coffee
  36. Pingback: kona coffee k cups
  37. Pingback: kona coffee
  38. Pingback: hualalai coffee
  39. Pingback: kona coffee
  40. Pingback: royal kona coffee
  41. Pingback: kona coffee
  42. Pingback: kona coffee
  43. Pingback: kktc üniversite
  44. Pingback: kona coffee
  45. Pingback: kona coffee
  46. Pingback: Sell used memory
  47. Pingback: Chicago Baler
  48. Pingback: Handmade molds
  49. Pingback: Pressemitteilung
  50. Pingback: cheap huawei mate8
  51. Pingback: Strom Angebot
  52. Pingback: mypsychicadvice
  53. Pingback: http://gp.se
  54. Pingback: Credit cards
  55. Pingback: Free Tablet Games
  56. Pingback: Numerology
  57. Pingback: outdoor end table
  58. Pingback: http://vlt.se/
  59. Pingback: fuck google
  60. Pingback: 他妈的谷歌
  61. Pingback: 他妈的谷歌
  62. Pingback: http://imdb.com
  63. Pingback: 他妈的谷歌
  64. Pingback: mobil porn
  65. Pingback: mobil porn
  66. Pingback: porno
  67. Pingback: 他妈的谷歌
  68. Pingback: iPhone
  69. Pingback: coffee from kona
  70. Pingback: 觀看色情肛門
  71. Pingback: gymshark
  72. Pingback: peaberry coffee
  73. Pingback: see this here
  74. Pingback: Web Wealth System
  75. Pingback: look at this site
  76. Pingback: Agen Bola
  77. Pingback: browse this site
  78. Pingback: Free stuff
  79. Pingback: computer repair
  80. Pingback: Sell event tickets
  81. Pingback: have a peek here
  82. Pingback: Piece Of Heaven
  83. Pingback: fukuidatumou
  84. Pingback: fukuidatumou
  85. Pingback: greek iptv
  86. Pingback: 色情管
  87. Pingback: Fun88 China
  88. Pingback: best coffee inc
  89. Pingback: hawaiian coffee
  90. Pingback: aquatic fish
  91. Pingback: in home care va
  92. Pingback: e cig liquid
  93. Pingback: more info here
  94. Pingback: escort orlando
  95. Pingback: oyunlar indir
  96. Pingback: multilingual seo
  97. Pingback: Make Money Online
  98. Pingback: QWEQEWQE
  99. Pingback: best gourmet
  100. Pingback: womens apparel
  101. Pingback: kona coffee bean
  102. Pingback: dry rock
  103. Pingback: download software
  104. Pingback: garden
  105. Pingback: descargar programs
  106. Pingback: Buy Tablet Binder
  107. Pingback: wildlife removal
  108. Pingback: code lookup
  109. Pingback: Ausgetrickst
  110. Pingback: Wirtschaftskanzlei
  111. Pingback: Marine Transport
  112. Pingback: Boîte à chocolat
  113. Pingback: Queen Hair Company
  114. Pingback: Led shower head
  115. Pingback: houses to buy
  116. Pingback: buy a home
  117. Pingback: Source
  118. Pingback: resume help
  119. Pingback: more info
  120. Pingback: outdoor sets
  121. Pingback: Auto loan
  122. Pingback: 牛混蛋
  123. Pingback: Enterprise
  124. Pingback: remote work
  125. Pingback: Business
  126. Pingback: travel
  127. Pingback: friv
  128. Pingback: Anime Online
  129. Pingback: other
  130. Pingback: penis enlargement
  131. Pingback: home garden towers
  132. Pingback: SATTA MATKA RESULT
  133. Pingback: cat supplies
  134. Pingback: MTN
  135. Pingback: Office 2016 pro
  136. Pingback: 福井脱毛
  137. Pingback: 밤알바
  138. Pingback: mp43gpfilm
  139. Pingback: Jaydani
  140. Pingback: sewing thread
  141. Pingback: BSM150GT120DN2
  142. Pingback: Theatre tickets
  143. Pingback: tampa bay magazine
  144. Pingback: hd1080hdkinoonline
  145. Pingback: matka
  146. Pingback: Turen
  147. Pingback: picccc
  148. Pingback: kona coffee beans
  149. Pingback: dab jars
  150. Pingback: custom paint
  151. Pingback: Home Security
  152. Pingback: funny pictures
  153. Pingback: saleforiphone
  154. Pingback: Home Automation
  155. Pingback: ICT
  156. Pingback: read more
  157. Pingback: details
  158. Pingback: ski chalets
  159. Pingback: Photography
  160. Pingback: robert
  161. Pingback: porn movies
  162. Pingback: black magic
  163. Pingback: istikhara online
  164. Pingback: Cheap Banners
  165. Pingback: skin care
  166. Pingback: satta matka
  167. Pingback: SATTA MATKA RESULT
  168. Pingback: Homepage
  169. Pingback: Lubitski
  170. Pingback: Play games online
  171. Pingback: Free Games
  172. Pingback: collegamento web
  173. Pingback: apple iPhone
  174. Pingback: project management
  175. Pingback: More information
  176. Pingback: lampamafussil
  177. Pingback: satta matka
  178. Pingback: click to read
  179. Pingback: youtube to mp3
  180. Pingback: Pinganillos
  181. Pingback: kinoklub
  182. Pingback: Click here
  183. Pingback: Dekorfliesen
  184. Pingback: personalised fans
  185. Pingback: Getting A Divorce
  186. Pingback: drama2016
  187. Pingback: Fenster
  188. Pingback: Turen
  189. Pingback: car
  190. Pingback: recipes
  191. Pingback: kala jadu
  192. Pingback: kala jadu
  193. Pingback: online istikhara
  194. Pingback: Best Companies
  195. Pingback: jpmsru
  196. Pingback: top2017bloomingme
  197. Pingback: sodick
  198. Pingback: adenosine
  199. Pingback: プラセンタ
  200. Pingback: プラセンタ
  201. Pingback: プラセンタ
  202. Pingback: プラセンタ
  203. Pingback: android tv box
  204. Pingback: what is the brain
  205. Pingback: wedding fans uk
  206. Pingback: phone case wood
  207. Pingback: gossip Lanka
  208. Pingback: me quipment
  209. Pingback: leer opinión
  210. Pingback: Car care
  211. Pingback: cats
  212. Pingback: anal sex
  213. Pingback: Garments supplier
  214. Pingback: IT Services
  215. Pingback: scam website
  216. Pingback: weird sex toys
  217. Pingback: shopping sex toy
  218. Pingback: anal sex toys
  219. Pingback: buttplug
  220. Pingback: anal beads
  221. Pingback: Best vibrator
  222. Pingback: Humans
  223. Pingback: Basic Human Rights
  224. Pingback: Be Happy
  225. Pingback: home design
  226. Pingback: cat food
  227. Pingback: David Miscavige
  228. Pingback: witches
  229. Pingback: cat food
  230. Pingback: g spot stimulator
  231. Pingback: luxury sex toys
  232. Pingback: sitemap
  233. Pingback: free ebooks
  234. Pingback: ebook
  235. Pingback: daddy dom stories
  236. Pingback: tamil sex story
  237. Pingback: 4sezon
  238. Pingback: TAXI DUBROVNIK
  239. Pingback: molodejka14seria
  240. Pingback: Hotels
  241. Pingback: dedicated server
  242. Pingback: molod17
  243. Pingback: trumpnews
  244. Pingback: Сталик
  245. Pingback: Hotwire Hotels
  246. Pingback: molodezhka4seria
  247. Pingback: Clarion Hotel
  248. Pingback: love quotes
  249. Pingback: mesothelioma law
  250. Pingback: men sex toys
  251. Pingback: molodejka
  252. Pingback: happiness
  253. Pingback: bunny sex toy
  254. Pingback: Rights
  255. Pingback: bdsm nipple clamps
  256. Pingback: UK website design
  257. Pingback: KODI
  258. Pingback: Letras de
  259. Pingback: Buy Seals Online
  260. Pingback: live auctions uk
  261. Pingback: no deposit slots
  262. Pingback: Drugs
  263. Pingback: Cloud solution
  264. Pingback: molodejka4sezon21s
  265. Pingback: link2016
  266. Pingback: xml181120167
  267. Pingback: Trenda Media
  268. Pingback: Sex Toy Reviews
  269. Pingback: free android games
  270. Pingback: istanbul escort
  271. Pingback: Yummy Mummy Outfit
  272. Pingback: kala jadoo
  273. Pingback: kala jadoo
  274. Pingback: molodejka4seria25
  275. Pingback: Couples Sex Toys
  276. Pingback: mdansby.com
  277. Pingback: Red Hearts
  278. Pingback: nighty
  279. Pingback: lastssadsafdsa
  280. Pingback: mdansby software
  281. Pingback: Vibrator
  282. Pingback: 福井歯医者
  283. Pingback: 福井歯医者
  284. Pingback: 福井歯医者
  285. Pingback: 福井歯医者
  286. Pingback: casino free slots
  287. Pingback: Mercedes-Benz
  288. Pingback: stalik hankishiev
  289. Pingback: DPS460EB A
  290. Pingback: Massager Vibrator
  291. Pingback: dadafdf3ddafdsa
  292. Pingback: dadafdf3ddafdsa
  293. Pingback: prehistorian
  294. Pingback: G-Spot Vibrators
  295. Pingback: best sex toys
  296. Pingback: Baler
  297. Pingback: flex vibrator
  298. Pingback: Penis Extension
  299. Pingback: Baler Manufacturer
  300. Pingback: classic coffee
  301. Pingback: current events
  302. Pingback: android phones
  303. Pingback: cialis
  304. Pingback: Tamworth
  305. Pingback: g-spot orgasm
  306. Pingback: butt plug
  307. Pingback: Surplus
  308. Pingback: Business for sale
  309. Pingback: pure kona
  310. Pingback: gastro
  311. Pingback: air jordan
  312. Pingback: Best Glass Dildo
  313. Pingback: Ejuice
  314. Pingback: finger vibrator
  315. Pingback: Fetish Restraints,
  316. Pingback: Get More Info
  317. Pingback: towing jobs
  318. Pingback: Consultant
  319. Pingback: RMUTT
  320. Pingback: N5K-C5548P-FA
  321. Pingback: adult shop
  322. Pingback: fleshlight videos
  323. Pingback: coupon discount
  324. Pingback: Webcam model jobs
  325. Pingback: athens
  326. Pingback: free casino slots
  327. Pingback: andrika
  328. Pingback: Nipple Clamps
  329. Pingback: best vibrator
  330. Pingback: tactical gear
  331. Pingback: tow trucks video
  332. Pingback: the best trucks
  333. Pingback: coffee belt kona
  334. Pingback: coffee belt kona
  335. Pingback: Anal Sex Lubricant
  336. Pingback: ohrayos
  337. Pingback: website hosting
  338. Pingback: workfromhomejobs
  339. Pingback: Waste Equip
  340. Pingback: Kristy May Young
  341. Pingback: iphone
  342. Pingback: delfi
  343. Pingback: personal training
  344. Pingback: 100% kona
  345. Pingback: 100% kona
  346. Pingback: this
  347. Pingback: webcam models
  348. Pingback: Toys Adults
  349. Pingback: toys for adults
  350. Pingback: air duct cleaning
  351. Pingback: Dryer
  352. Pingback: sex sling
  353. Pingback: kalatburti
  354. Pingback: jack rabbit toy
  355. Pingback: bitcoin faucet
  356. Pingback: buy penis ring
  357. Pingback: toronto rap
  358. Pingback: webcam models
  359. Pingback: anal toy
  360. Pingback: pure kona coffee
  361. Pingback: bangla choti
  362. Pingback: buy kona coffee
  363. Pingback: xbox games
  364. Pingback: best kona
  365. Pingback: pure kona
  366. Pingback: NasstoysNewYork
  367. Pingback: prostate
  368. Pingback: ciao amigos
  369. Pingback: toronto homes
  370. Pingback: maxie solters
  371. Pingback: Discover
  372. Pingback: Birdman Mixtapes
  373. Pingback: g-spot vibrator
  374. Pingback: Fabolous Mixtapes
  375. Pingback: Fetty Wap Mixtapes
  376. Pingback: group chat app
  377. Pingback: Male Enhancement
  378. Pingback: Dealer Advertising
  379. Pingback: vibrators
  380. Pingback: 롤대리랭
  381. Pingback: sex stories
  382. Pingback: Yoga
  383. Pingback: dolly towing
  384. Pingback: Greg Thmomson
  385. Pingback: Sex Stories
  386. Pingback: best sex toys ever
  387. Pingback: sexy underwear
  388. Pingback: anal toy
  389. Pingback: sex lubricant
  390. Pingback: sex restraints
  391. Pingback: Wall decals
  392. Pingback: adult toy
  393. Pingback: impresoras
  394. Pingback: meet 50+ singles
  395. Pingback: bali silver
  396. Pingback: piezas suzuki
  397. Pingback: website hosting
  398. Pingback: free pic upload
  399. Pingback: Best Cock Ring
  400. Pingback: pronájem plošiny
  401. Pingback: Ford F 350
  402. Pingback: lovedolls
  403. Pingback: anal plus fun men
  404. Pingback: white morkie dog
  405. Pingback: vibrator wand
  406. Pingback: film streaming
  407. Pingback: gmail log in
  408. Pingback: best male sex toy
  409. Pingback: oral surgery
  410. Pingback: tablet repair
  411. Pingback: 스포츠닥터스
  412. Pingback: pure kona coffee
  413. Pingback: 스포츠닥터스
  414. Pingback: best kona coffee
  415. Pingback: Triple Stimulation
  416. Pingback: Berber carpet
  417. Pingback: ways to make money
  418. Pingback: How to Buy Online
  419. Pingback: usuwanie dpf
  420. Pingback: Buy Pocket Pussy
  421. Pingback: adamandeve
  422. Pingback: tech forum
  423. Pingback: CISO Terbaik 2017
  424. Pingback: young beagle
  425. Pingback: chug
  426. Pingback: pdr training
  427. Pingback: nesaporn
  428. Pingback: the best cock ring
  429. Pingback: Mack LR Series
  430. Pingback: movie malayalam
  431. Pingback: PHP Music website
  432. Pingback: Acoustic
  433. Pingback: amada edm filter
  434. Pingback: cat kennels
  435. Pingback: wRfqnI8IRxk
  436. Pingback: dabs
  437. Pingback: SAI
  438. Pingback: Double Vibrator
  439. Pingback: free app download
  440. Pingback: app for laptop
  441. Pingback: Boudoir
  442. Pingback: cars
  443. Pingback: sexshop bh
  444. Pingback: 100% kona lion
  445. Pingback: Fixed ROM
  446. Pingback: buy kona coffee
  447. Pingback: pure kona beans
  448. Pingback: strapon
  449. Pingback: clit vibrator
  450. Pingback: miniclip.com
  451. Pingback: Best Butt Plugs
  452. Pingback: Chaga Pilz
  453. Pingback: Masturbator Toy
  454. Pingback: nike air jordan
  455. Pingback: Sears
  456. Pingback: strap on toys
  457. Pingback: mulvadi pure kona
  458. Pingback: 木瓜
  459. Pingback: Rabbit Vibrators
  460. Pingback: why do wives cheat
  461. Pingback: telefonsex
  462. Pingback: erotik
  463. Pingback: peru travel
  464. Pingback: all of craigslist
  465. Pingback: telefonsex mit cam
  466. Pingback: cash loans
  467. Pingback: gym diet
  468. Pingback: leather dress
  469. Pingback: telefonsex cam
  470. Pingback: Good Vibes Toys
  471. Pingback: wet rabbit
  472. Pingback: Phil Doe
  473. Pingback: masturbation
  474. Pingback: climax the series
  475. Pingback: sex toys
  476. Pingback: vagina
  477. Pingback: szybki kredyt
  478. Pingback: sex therapist
  479. Pingback: vape near me
  480. Pingback: designer
  481. Pingback: forex live signal
  482. Pingback: amazon wish list
  483. Pingback: camere spion
  484. Pingback: persian setar
  485. Pingback: cheapest smm panel
  486. Pingback: g spot vibrator
  487. Pingback: feather sex toy
  488. Pingback: rabbit vibe
  489. Pingback: destiny
  490. Pingback: g-spot vibrator
  491. Pingback: suction cup dong
  492. Pingback: porn stories
  493. Pingback: cock ring
  494. Pingback: sd card recovery
  495. Pingback: adam and eve
  496. Pingback: Freshest food
  497. Pingback: Pinganillos
  498. Pingback: Pinganillo
  499. Pingback: Arduino sensors
  500. Pingback: Pinganillo
  501. Pingback: here
  502. Pingback: pet travel
  503. Pingback: 福井脱毛
  504. Pingback: 福井脱毛
  505. Pingback: 福井脱毛
  506. Pingback: 福井脱毛
  507. Pingback: bhutan weed
  508. Pingback: c fashion magazine
  509. Pingback: fashion week milan
  510. Pingback: kinokradserial
  511. Pingback: brandt 0 18 42
  512. Pingback: online beats
  513. Pingback: ladies clothes
  514. Pingback: fashion-element-7
  515. Pingback: buy kona
  516. Pingback: park ridge
  517. Pingback: california
  518. Pingback: pure kona
  519. Pingback: homer p figg
  520. Pingback: sex diary
  521. Pingback: home page
  522. Pingback: cool stuff
  523. Pingback: Beads
  524. Pingback: dealing with std
  525. Pingback: dating tips
  526. Pingback: Sex Toy Storage
  527. Pingback: Buy Dildo
  528. Pingback: best kona
  529. Pingback: pure kona
  530. Pingback: TensorFlow
  531. Pingback: fishing redding
  532. Pingback: dentist
  533. Pingback: legian
  534. Pingback: Strap On Dildo
  535. Pingback: Birthday images
  536. Pingback: world news
  537. Pingback: wedding tips diy
  538. Pingback: annabelle knight
  539. Pingback: women clothing
  540. Pingback: rehab clinic
  541. Pingback: women's health
  542. Pingback: retro 13 jordans
  543. Pingback: cock rings
  544. Pingback: 100% pure kona
  545. Pingback: masturbation
  546. Pingback: Dry Skin Treatment
  547. Pingback: Zastava Yugo
  548. Pingback: usa email lists
  549. Pingback: home page
  550. Pingback: Make Money Online
  551. Pingback: wall bed
  552. Pingback: Voodoo
  553. Pingback: sport bra
  554. Pingback: unique antiques
  555. Pingback: vibrator for her
  556. Pingback: butterfly sex toys
  557. Pingback: sex toys cleaner
  558. Pingback: sex toy review
  559. Pingback: adam eve toys
  560. Pingback: bullet sex toy
  561. Pingback: dvd專賣店
  562. Pingback: ean code
  563. Pingback: 100% pure kona
  564. Pingback: Viagra kaufen
  565. Pingback: Gadgets
  566. Pingback: free web hosting
  567. Pingback: sex toys
  568. Pingback: female vibrator
  569. Pingback: business terms
  570. Pingback: rabbit vibrator
  571. Pingback: nipple orgasm
  572. Pingback: 12win singapore
  573. Pingback: h real estate logo
  574. Pingback: full cotton sheets
  575. Pingback: marketing s&p
  576. Pingback: h and v industry
  577. Pingback: nexus 7 shopping
  578. Pingback: 3 education cess
  579. Pingback: 2 fashion sisters
  580. Pingback: food quiz
  581. Pingback: vibrator
  582. Pingback: health help
  583. Pingback: 3 news articles
  584. Pingback: j healthc qual
  585. Pingback: vibe
  586. Pingback: bangal ka jadu
  587. Pingback: forex trading
  588. Pingback: but plug review
  589. Pingback: all forex brokers
  590. Pingback: Quick News Updates
  591. Pingback: Hot Hot Hot Site
  592. Pingback: g spot vibe
  593. Pingback: couples sex toy
  594. Pingback: auto detailing
  595. Pingback: serial 2017
  596. Pingback: nylon cutter head
  597. Pingback: label besi
  598. Pingback: butt plug
  599. Pingback: gay toys
  600. Pingback: best gay sex toys
  601. Pingback: anal toy
  602. Pingback: Female Sex Toys
  603. Pingback: vibrating dildo
  604. Pingback: Candles
  605. Pingback: wet wabbit
  606. Pingback: bandiera italiana
  607. Pingback: смотреть
  608. Pingback: kinogo
  609. Pingback: kala jadu
  610. Pingback: Butterfly Sex Toy
  611. Pingback: Arousal Cream
  612. Pingback: Bondage Restraint
  613. Pingback: dick extension
  614. Pingback: personal lubricant
  615. Pingback: Sripatum
  616. Pingback: feather tickler
  617. Pingback: robert

Leave a Reply