Figure 10.

CMA regulates lipid droplet accumulation through ACLY and ACSS2. (A) NIH3T3 cells were stained with BOPIDY (lipid droplets) and Hoechst (DNA), 96 h after transfection with the indicated siRNAs. Scale bars: 10 μm. (B) Western blot validation for knockdown efficiency for siRNAs against Lamp2a, Acly, and Acss2. (C) Quantification of BOPIDY staining intensity across whole fields of confluent cells, imaged with identical exposure times, with no post-acquisition image adjustment. Four fields for each of four experimental repeats are pooled, for a total of 16 data points for each group. (D) Western blot validation for knockdown efficiency for siRNAs against Lamp2a and Fasn. (E) NIH3T3 cells were stained with BOPIDY and Hoechst 96 h after transfection with the indicated siRNAs. Scale bars: 20 μm. (F) Quantification of BOPIDY staining, using the same methods as (C). p-values are replaced with asterisks to save space: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. For (D) results of a 2-way ANOVA are shown in the table under the graph.