FIGURE 5:

Effects of lysosome fusion inhibition on the CtxB-staining fluorescent patches and ceramide. (A) Percentage changes in positive cells costained by Al488-CtxB and anticeramide antibody in FasL-stimulated CAECs pretreated with lysosome fusion inhibitor vacuolin-1 (10 μM), ASM inhibitor Ami (20 μM), ASM siRNA, V1 H⁺-ATPase siRNA, and its inhibitor Baf (100 nM). (n = 6, p < 0.05 vs. control; ^#p < 0.05 vs. only FasL-treated group). (B) Distribution and localization of H⁺-ATPase as a MR redox platform on the membrane in CAECs treated with FasL alone or with FasL after pretreatment of V1 H⁺-ATPase siRNA. Fractions 3–5 were designated as MRs, as indicated by the marker protein flotillin-1. The blot pattern for H⁺-ATPase represents four individual experiments. Na⁺/K⁺-ATPase could also be detected in membrane raft fractions. However, no marked increase in the Na⁺/K⁺-ATPase protein in MR microdomains was observed in FasL-treated CAECs.