Fig. 3

KDS-5104 increases lysosomal biogenesis in a PPARα dependent manner. (a) Representative LAMP1 immunofluorescence images of vehicle or KDS-5104 treated BV2 cells with or without GW6471 pretreatment. (b) Quantification of LAMP1 fluorescent intensity (n = 6/condition). (c) Representative images of LAMP1 (green) and DAPI (white) staining and corresponding 3D renderings of primary WT and PPARαKO microglia cultures treated with vehicle or 10 µM KDS-5104 for 8 h. (d) Quantification of lysosome size, number of lysosomes per cell and distance of lysosomes from the nucleus. (e) Representative images of lysotracker positive lysosomes and 3D renderings of primary WT and PPARαKO microglial cultures treated with vehicle or 10 µM KDS-5104 for 8 h. (f) Quantification of lysotracker fluorescence intensity and number of lysotracker + lysosomes per cell. Approximately 25 cells obtained from a minimum of 3 wells per group were analyzed. AU: artificial unit. Scale bars: 200 μm in (a) and 100 μm in (c, e). For all panels, data are presented as mean ± SEM. ns: non-significant, *p < 0.05, **p < 0.01, ****p < 0.0001. One way ANOVA with Tukey’s multiple comparisons tests as the post hoc analysis