Figure 4: ACF7 regulates wound repair and tight junction dynamics of intestinal epithelial cells in vivo.
(a) The representative sections of the intestines from the WT and ACF7 cKO mice at 4 and 24 h post BrdU pulse show that the migration of BrdU-positive cells from the crypt is inhibited by deletion of ACF7. Scale bar, 50 μm. (b) Quantification of BrdU-positive cell migration in vivo, indicating significantly decreased migration in the ACF7 cKO mice compared with the WT mice (n=10, ***P<0.001, Student’s t-test). Sample size n>9 (three independent tests with different animals, and three technical replicates each). (c) Oligocellular wounds were introduced to colonic epithelium by laser ablation in vil-mRFP-ZO1 transgenic mice with ACF7 cKO or WT background. Would healing in vivo is monitored by intravital imaging with multiphoton microscopy. Scale bar, 50 μm. (d) Wound healing in vivo was quantified by measuring the percentage of wound closure over time with ImageJ. Note WT heals significantly faster than cKO mice. Sample size n=3 (three independent tests on different animals), mean±s.d. (e) FRAP analysis was carried out to determine the dynamics of tight junctions. Montages show fluorescence recovery of mRFP-ZO1 in vivo. Scale bar, 50 μm. (f) Box and whisker plotting of T1/2 (half time) for FRAP analysis in WT and cKO colon epithelium. Deletion of ACF7 significantly increases T1/2 in vivo (P<0.01, Student’s t-test). Sample size n>9 (three independent tests, and three technical replicates each).
