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Fig. 5. Fate of claudin-1 and claudin-2 during apoptosis of epithelial cells. (A) MDCK and H184A1 cell lysates (5 µg) were analyzed at different time points after induction of apoptosis with staurosporine by western blotting with polyclonal rabbit anti-claudin-1 antibody. (B) Apoptosis was induced in MDCK cells by treatment with TNF- ${alpha}$ for 24 hours (lane 1, 0 hours; lane 2, adherent cells 24 hours; lane 3, floating cells 24 hours). (C) HT-29/B6 cell lysates (5 µg) were analyzed at different time points after induction of apoptosis with staurosporine by western blotting with polyclonal anti-claudin-1 or anti-claudin-2 antibodies, respectively. On both western blots identical amounts of protein from the same cell lysate were analyzed. Western blots show representative data from four independent experiments. (D) Ltk^- cells were transiently transfected with full-length claudin-2-FLAG₃. Apoptosis was induced with 1 µM staurosporine for 6 hours. Western blots of cell lysates (40 µg) were analyzed with monoclonal anti-FLAG M2 antibody. Lane 1, Ltk^- cells transfected with empty vector; lane 2, control Ltk^- cells transfected with claudin-2-FLAG₃; lane 3, Ltk^- cells transfected with claudin-2-FLAG₃ 6 hours after induction of apoptosis with staurosporine. Western blots are representatives of at least three independent experiments.

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