Fig. 4: ACOT12 may be involved in cholesterol trafficking by interacting with VPS33A.
From: Loss of Acot12 contributes to NAFLD independent of lipolysis of adipose tissue
a Representative filipin staining of Acot12+/+ and Acot12−/− primary hepatocytes. Scale bars, 25 μm. b Representative immunocytochemical staining for ACOT12 in the NCTC clone 1469 normal mouse liver cell line with or without 50 μg/ml water-soluble cholesterol treatment. Scale bars, 100 μm. Fluorescence intensity was measured in five randomly collected images by ImageJ and is visualized as a bar graph (right). c Treatment of BODIPY cholesterol in Acot12+/+ and Acot12−/− primary hepatocytes. Scale bars, 20 μm. d Diagram of the location and function of ACOT12-interacting proteins as assessed by BioID. e Merged image of RFP-ACOT12 and EGFP-VPS33A. Scale bars, 20 μm. f Representative merged image of EGFP-fused VPS33A (green, EGFP-VPS33A) and LysoTracker staining (red). Scale bars, 20 μm. g Merged and magnified images (a’ and b’) of TagRFP-fused ACOT12 (red, RFP-ACOT12) and BODIPY cholesterol (green, BD-Chol). Scale bars, 100 μm. h Representative image of LysoTracker and BD-Chol in Acot12+/+ and Acot12−/− primary hepatocytes. Nuclei were stained by Hoechst 33342. Scale bars, 50 μm. Fluorescence intensity was measured in ten randomly collected images by ImageJ and is visualized as a bar graph (right). All data are presented as the mean ± SD. Statistical differences between the two groups were determined using an unpaired two-tailed Student’s t test. **P < 0.01.
