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. 2017 Nov 17;6:e30543. doi: 10.7554/eLife.30543

Figure 3. Loss of Bax and Bak1 maintains lysosomal integrity.

Figure 3.

(A) Transmission EM images (30,000x) of WT and DKO MEFs subjected to serum starvation for the specified time points. Autolysosomes and translucent vesicles are noticeable in the serum-starved WT MEFs but not in the DKO MEFs. Scale bars = 1 μm. (B) Confocal images of WT and DKO MEFs subjected to serum starvation for the indicated times and labeled with Rhodamine 123 (green) to measure mitochondrial membrane potential and LysoTracker red (red) to label lysosomes. Scale bars = 5 μm. (C) Time course of LysoTracker red fluorescence intensity in serum starved WT vs. DKO MEFs for the indicated time points. (D) Representative trace for LysoTracker red fluorescence in WT and DKO MEFs serum-starved for 12 hr and treated with or without bafilomycin A1. Fluorescent intensity was continually measured for 40 min following the bafilomycin A1 addition. (E) Confocal images of WT and DKO MEFs treated the same as in panel ‘D’. Scale bars = 10 μm. (F) Graph of cytosolic pH in WT and DKO MEFs in control serum-replete conditions (black) or serum-starved (red) conditions. (G) Confocal images of active cathepsin B (red) in WT and DKO MEFs in control or serum-starved state. Scale bars = 5 μm. All assays are an average, or are representative of three independent experiments. The error bars represent the standard error of the mean. *p<0.05 vs 0 serum-replete. Statistical significance was determined by student's t-test.