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. 2020 Jul 29;42:101055. doi: 10.1016/j.molmet.2020.101055

Figure 6.

Figure 6

Arf1 is required for cholesterol transport into mitochondria. (A) Confocal imaging analysis depicting the mitochondrial cholesterol transport in HeLa vector control (VC) and Arf1 knockout (Arf1-KO) cells. Cells were starved in KRPH buffer for 30 min and then treated with 10 μM of CholEsteryl BODIPY™ 542/563 C11 for 10 min. (B) Confocal imaging analysis depicting the mitochondrial cholesterol transport in C2C12 cells treated with 10 μM of Exo-2 for 2 h followed by incubation with 10 μM of CholEsteryl BODIPY™ 542/563 C11 for 10 min. Nuclei were stained with Hoechst 33342, and mitochondria were stained with Mitotracker-Green. (C–E) Total cholesterol level (C) and cholesterol levels in pure mitochondria (D) and MAM (E) in HeLa VC and Arf1 KO cells. Cells were treated with MCD (-CHOL), or pemeabilized by digitonin after MCD treatment and re-incubated with 50 μM of cholesterol (+CHOL). n = 3. ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001 by two-way ANOVA.