FIGURE 3:

ARL8 KO increases lysosomal cholesterol and decreases LDs. (A) Halo tag was fused to endogenous ARL8a and ARL8b isoforms by CRISPR-Cas9 in WT or myrlysin-KO HeLa cells. Halo tag and lysosomes were visualized by applying Halo dye and LysoTracker in live-cell imaging experiments, respectively. Cells as indicated were fixed and then stained with filipin (B) or stained with BODIPY (D). Fluorescence signals were observed by confocal microscopy, quantified, and normalized to the signals in WT cells. For filipin quantification, n = 105, 111, 131, and 127. For BODIPY quantification, n = 119, 88, 77, and 60. (E, F) Bar graphs are presented as mean ± SD; p values were determined using Student’s t test. ***p < 0.001 (vs. WT). Scale bars, 5 μm.