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Figure 1


Fig. 1. PAR6{Delta}N promotes apoptosis in MDCK cystogenesis. (A,B) Wild-type MDCK and Myc-tagged PAR6{Delta}N-expressing stable cell lines grown on collagen for 7 days were fixed, permeabilized and immunostained for Myc (green), beta-catenin (red) and ZO-1 (white) (A) and for Myc (green), actin (red) and cleaved caspase-3 (white) (B). White arrow and arrowhead indicate dead cells in A. Nuclei are shown as blue in all figures. (C) The levels of apoptosis were measured using a cell death ELISA. Results are the mean±s.d. of three experiments. (D) Wild-type MDCK and PAR6{Delta}N lysates isolated from cysts were prepared at different time points (day 5, 7 and 13) and then analyzed for activation of caspase-3 using cleaved caspase-3 and GAPDH antibodies (loading control). (E) PAR6{Delta}N cysts were treated with either a carrier (DMSO) or caspase inhibitor, zVAD-fmk (50 µM) three times at day 0, 4 and 6 and immunostained for cleaved caspase-3 (red) at day 7, as described in A. Bars, 10 µm.





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