Fig. 3.

Silencing of MST4 decreases lipid accumulation in IHHs. IHHs were transfected with MST4 siRNA or NTC siRNA and challenged with OA for 48 hours. (A) Schematic illustration of the study design. (B,C) MST4 mRNA (B) and protein (C) abundance. Protein levels were analyzed by densitometry; representative western blots are shown with GAPDH used as a loading control. (D) Representative immunofluorescence images of cells double‐stained with antibodies for MST4 (green) and ADRP (red); nuclei stained with DAPI (blue). (E,F) Representative images of cells stained with Bodipy (green) or MitoTracker Red (red), or processed for immunofluorescence with anti‐PEX5 (red) or anti‐PMP70 (red) antibodies; nuclei stained with DAPI (blue). Quantification of the staining. (G) Oxidation of radiolabeled palmitate. (H) Secretion of [³H]TAG into the media. (I) Fatty acid uptake rate. (J) TAG synthesis from [¹⁴C]‐labeled glucose. (K) Glycogen levels. (L) Cell viability was assessed using resazurin. In (D) and (F), the scale bars represent 10 µm and 25 µm, respectively; in (E), the scale bars at the left and right represent 25 µm and 10 µm, respectively. Data are presented as mean ± SEM from 4‐10 wells per group. Abbreviation: Transf, transfection. *P < 0.05, **P < 0.01, ***P < 0.001.