Fig. 2. Olanzapine induces lipid droplets accumulation, lysosome enlargment and cholesterol entrapment.

A HepG2 cells serum-starved overnight and pre-treated with DMEM low glucose 20% FBS for 3 h, were incubated with the indicated compounds for 24 h. Cells were fixed and incubated with a solution of Oil-Red-O 0.2% w/v in 40% v/v isopropanol for 30 min and counterstained with DAPI. The lipid droplets fluorescence (n = 65 cells), the percentage of cell area covered by lipid droplets (n = 40 cells), and the lipid droplets diameter (n = 250 vesicles) were quantified and reported in the graph (one-way ANOVA followed by Dunnett’s multiple comparison test; *vs unt). Scale bar =10 µm. B Treated cells were fixed and incubated with 100 µg/ml Filipin III (blue) and anti LAMP1 antibody (green). Several parameters were quantified from panel B and reported in the graphs: C the diameter of Filipin III positive accumuli (n = 400 vesicles), and the percentage of cell area covered by Filipin III accumuli (n = 45 cells), D lysosomal diameter (n = 400 vesicles) and lysosomal number (n = 45 cells) (one-way ANOVA followed by Dunnett’s multiple comparison test). Scale bar =10 µm.