Figure 1
BET bromodomain inhibition does not alter the gross structure of the small intestine.
(A) Western blot analysis of Brd2, Brd3 and Brd4 in whole cell lysates from preparations of crypts and villi from the jejunum. The predominant expression of Villin and PCNA in the villi and crypts, respectively, served as controls for the proper enrichment of these compartments. β-actin served as a loading control. Results are representative of three independent experiments. (B) RNA was isolated from purified villi and crypts from the jejunum of mice treated with vehicle control or BETi for 24 hours and the expression of PHF15 was evaluated. The average (±std. dev.) relative expression from three mice is shown. Differences in the expression that were statistically significant (p < 0.05) are indicated by an asterisk. (C) H&E images of the small intestine of mice after 14 days of treatment with BETi or vehicle control. The villus height (avg. ± std. dev.) measured from 3 control and BETi treated mice are shown. (D) Intestinal alkaline phosphatase activity was measured using p-nitrophenyl phosphate (pNPP) as a substrate with cell lysates from the small intestine of mice harvested after 14 days of treatment with BETi or vehicle control. The averages (±std. dev.) from three mice are shown. The differences in pNPP activity were not significant (n.s., p = 0.12). (E) Representative images of immunohistochemistry for BrdU incorporation in the crypts of the small intestine in mice after 7 and 14 days of treatment with BETi or vehicle control. The average (±std. dev.) number of BrdU positive cells per crypt is shown. (F) Intestinal permeability was determined serum levels of FITC-dextran after gastric gavage in mice after 14 days of treatment with vehicle control or BETi. The averages (±std. dev.) from 3 mice are shown.
