Figure 3.

TG/LDs Accumulate upon the Inhibition of Fatty Acid β-Oxidation under Glucose-Free Conditions, Leading to Increased GBM Cell Death

(A and B) Representative fluorescence images of LDs stained with BODIPY 493/503 (green)/Hoechst 33342 (blue) (A). TLC analysis of total lipids (B) in U87 (8 h), U251, and T98 cells (4 h) after treatment with/without ETO (100 μM) in the absence or presence of glucose (25 mM). LDs/cell (mean ± SEM, n = 30) and TG levels (mean ± SD, n = 3) were quantified by ImageJ and normalized to the control cells.

(C) Cell death was determined in U87 (8 h), U251, and T98 cells (3 h) after treatment with/without ETO (100 μM) in the absence or presence of glucose (25 mM) by trypan blue staining (mean ± SD, n = 3).

(D–G) U87 cells were transfected with CPT1A shRNA-expressing lentivirus for 24 h and then cultured in glucose-free or 25 mM glucose medium for 8 h. Cells were analyzed by western blotting (D), observed by confocal microcopy after BODIPY 493/503 and Hoechst 33342 staining (E), or analyzed by TLC for lipid levels (F). LDs/cell (mean ± SEM, n = 30) or relative TG levels (mean ± SD, n = 3) were determined by ImageJ. Cell death was determined after trypan blue staining (mean ± SD, n = 3) (G). FA, fatty acids.

Significance was determined by one-way ANOVA. ∗∗p < 0.01, ∗p < 0.05. Please also see Figure S2.