FIG 8.

Ectopically expressed uc.173 prevents miR-29b-induced epithelial barrier dysfunction. (A) Levels of miR-29b (a) and uc.173 (b) 48 h after cells were either transfected with pre-miR-29b alone or cotransfected with pre-miR-29b and a uc.173 expression vector. O/E, overexpression. Values are relative to those with a scrambled control oligomer and are means ± SEM of data from triplicate experiments. Asterisks indicate significant differences (P < 0.05) from results for cells transfected with the scrambled oligomer. (B) Effect of increasing uc.173 levels on the association of miR-29b with CLDN1 mRNA. Twenty-four hours after cells were transfected with a uc.173 expression vector, they were transfected with biotin-labeled miR-29b (Bio-miR-29b). The levels of CLDN1 mRNA in the materials pulled down by miR-29b were examined 24 h thereafter. The asterisk and plus sign indicate significant differences (P < 0.05) from the results for cells transfected with a scrambled oligomer or for cells transfected with Bio-miR-29b and a control vector, respectively. (C) Representative immunoblots of claudin-1, occludin, and E-cadherin proteins in cells treated as described for panel A. (D) Changes in TEER (left) and FITC-dextran paracellular permeability (right) in cells treated as described for panel A. Values are means ± SEM of data from six samples. The asterisks and plus sign indicate significant differences (P < 0.05) from results for cells transfected with a scrambled oligomer or with pre-miR-29b alone, respectively.