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. 2022 Feb 16;298(3):101740. doi: 10.1016/j.jbc.2022.101740

Figure 2.

Figure 2

Loss of INPP5A increases the lysosomal volume fraction and lysosomal proteolytic capacity.A, quantification of the lysosomal volume fraction normalized to the cytoplasm of HeLa cells treated with control (SCR) or INPP5A siRNA (5AKD). Unpaired t test. p = 0.0408, t = 2.106, and df = 45. B, representative electron micrographs of HeLa cells treated with control (SCR) or INPP5A siRNA (5AKD). Note the accumulation of degradative lysosomal organelles in 5AKD cells. The scale bar represents 1 μm. C, zoomed area of white boxes in B. The scale bar represents 250 nm. D, quantification of cathepsin L Magic Red activity in HeLa cells treated with control (SCR) or INPP5A siRNA (5AKD) one-sample t test. p = 0.0155, t = 7.95, and df = 2. E, representative confocal images of D. Blue, DAPI-stained nuclei. The scale bar represents 10 μm. F, representative immunoblot analysis of HeLa cells treated with control (SCR) or INPP5A siRNA (5AKD). Immunoblots were decorated with antibodies against LC3 and p62. G, quantification of LC3 puncta in HeLa cells treated with control (SCR) or INPP5A siRNA (5AKD) one-sample t test. p = 0.0110, t = 9.440, and df = 2. H, representative confocal images of G. Blue, DAPI-stained nuclei. The scale bar represents 10 μm. Data represent n = 23 cells for SCR and n = 24 cells for 5AKD in EM; three independent experiments for cathepsin L Magic Red and LC3 staining. DAPI, 4′,6-diamidino-2-phenylindole; INPP5A, inositol polyphosphate-5-phosphatase A.