Fig. 6

Silencing of TAOK3 in vitro suppresses lipid accumulation and oxidative stress in cultured mouse hepatocytes without inducing a compensation response by related STE20-type kinases. Primary hepatocytes isolated from wild-type mice were transfected with mouse Taok3 siRNA or NTC siRNA and cultured with oleate supplementation. A TAOK3 protein abundance assessed by Western blot. Protein levels analyzed by densitometry; representative Western blots are shown with glyceraldehyde-3-phosphate dehydrogenase (GAPDH) used as a loading control. B, C Representative images of mouse hepatocytes stained with Bodipy 493/503 (green; B) or DHE (red; C); nuclei stained with DAPI (blue). The scale bars represent 25 µm. Quantification of the staining. (D) Relative mRNA expression of selected STE20 kinases assessed by qRT-PCR. Data are mean ± SEM from 3 (A) or 6 (B–D) wells per group. Statistical significance between the groups was evaluated using the unpaired 2-tailed Student’s t-test. *P ≤ 0.05, ***P < 0.001