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. 2015 Aug 3;210(3):435–450. doi: 10.1083/jcb.201501002

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Figure 7. — TFEB KO cells do not exhibit a defect in mitophagy. (A) WT and TFEB KO cells stably expressing mCherry-Parkin were treated with DMSO or O/A for up to 24 h, lysed, and immunoblotted. (B) Cells from A were treated with DMSO or O/A for 24 h. Fixed cells were immunostained for DNA and TOM20 and analyzed by immunofluorescence. Bars, 20 µm. (C) WT and TFEB KO cells stably expressing YFP-Parkin and mt-mKeima were treated with DMSO or O/A for 18 h and subjected to FACS analysis. Plots are representative of n = 2 experiments. (D) Quantification of MTCO2 levels in A. MTCO2 expression was normalized to actin and expressed relative to 0-h treatment. Data are means ± SD (n = 3). (E) Quantification of anti-DNA stain in B. Z stacks of five to seven fields per condition (20–30 cells/field) were analyzed per experiment (n = 3). Data are means ± SD. (F) Proportion of cells in the lower gate in C. Data are means ± range (n = 2 experiments). (G and H) WT and TFEB KO cells lacking endogenous Parkin (G) or stably expressing mCherry-Parkin (H) were treated with DMSO or O/A for 18 h, lysed, and immunoblotted. Three independent experiments (a–c) are shown.