Fig. 1: Cholesterol depletion promotes autophagy initiation. | Nature Communications

Fig. 1: Cholesterol depletion promotes autophagy initiation.

From: The cholesterol transport protein GRAMD1C regulates autophagy initiation and mitochondrial bioenergetics

Fig. 1

a U2OS cells were treated with 2.5 mM methyl-β cyclodextrin (MBCD) in DMEM or in EBSS supplemented ± 10 nM Bafilomycin A1 (BafA1) for 1 h. b Quantification of LC3B-II band intensity relative to actin and normalized to DMEM control. Error bars = SEM. Significance was determined using one-way ANOVA followed by Tukey’s comparisons test from n = 3 independent experiments. P value <0.001 and 0.004. c U2OS cells were treated with 2.5 mM MBCD in DMEM or in EBSS ± 10 nM BafA1 for 1 h. Scale bar = 20 µm. d Quantification of LC3B puncta number per cell normalized to the DMEM control. Error bars = SEM. Significance was determined using one-way ANOVA followed by Tukey’s comparisons test from n = 4 independent experiments, >500 cells per condition. P value = 0.001 and 0.006. e U2OS cells were treated with 10 µM atorvastatin (ATV) for 48 h, before amino acid starvation in EBSS 1 h ±10 nM BafA1. f Quantification of LC3B-II band intensity relative to actin and normalized to DMEM control. Error bars = SEM. Significance was determined using one-way ANOVA followed by Tukey’s comparisons test from n = 3 independent experiments. P value = 0.0123. g U2OS cells were treated with 2.5 mM MBCD in DMEM or EBSS. Scale bar = 20 µm. h, i ATG13 (h) and ATG16L1 (i) puncta number per cell were normalized to the DMEM control. Error bars = SEM. Significance was determined using two-way ANOVA followed by Sidak’s comparisons test from n = 3 independent experiments, >500 cells per condition. P value (ATG13) < 0.0001, <0.0001 and P value (ATG16L1) = 0.0008 and P value <0.0001. j U2OS cells were treated with 10 µM ATV for 48 h, before amino acid starvation in EBSS 1 h Scale bar = 20 µm. k ATG13 puncta number per cell were normalized to the DMEM control. Error bars = SEM. Significance was determined using two-way ANOVA followed by Sidak’s comparisons test from n = 3 independent experiments, >500 cells per condition. P value = 0.0437 and 0.0005. Source data are provided as a Source Data file.

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