(A) Effects of inhibitors on LY accumulation by cells growing in BMM, measured after 5 hrs in 500 μg*ml−1 LY. Ordinary one-way ANOVA compared experimental to control values; ns, not significant; ****, p<0.0001.
(B) Effects of inhibitors on LY efflux from lysosomes. Cells were pulsed 2 hr in BMM containing 500 μg*ml−1 LY, then were washed and chased 3 hr in BMM containing the indicated inhibitors. Dishes were then washed and lysed to measure remaining cell-associated LY. Higher values for retention indicate inhibition of recycling. J/B decreased LY retention and SAR-405, bafilomycin A1, Torin-1 and rapamycin increased retention. One-way ANOVA compared to control values; ns, not significant; ***, p<0.001, ****, p<0.0001. (C) Inhibitors were added to cells growing in BMM at 18 hr after the beginning of cell culture (arrow). Cell proliferation in BMM was inhibited by J/B, SAR-405, bafilomycin A1, rapamycin and Torin-1.