Fig. 3: PIN1 negatively regulates MATα1 mitochondrial targeting.

Western blots of MATα1 and PIN1 in AML-12 and HepG2 cells after PIN1 silencing (a) and overexpression (b). Western blots and densitometry analyses of cytosolic and mitochondrial MATα1 and PIN1 in AML-12 and HepG2 cells after PIN1 silencing (c and e, AML-12: n = 4 independent experiments, p = 0.033 and HepG2: n = 3 independent experiments, p = 0.022) and overexpression (d and f, AML-12: n = 3 independent experiments, p = 0.003 and HepG2: n = 3 independent experiments, p = 0.009). AML-12 are shown in dark blue and HepG2 in light blue. g Western blot and h densitometry analyses of total and mitochondrial MATα1 in AML-12 cells after PIN1 silencing and ethanol treatment (n = 3 independent experiments, p = 0.006 for total and p = 0.005 for mitochondrial MATα1 SC ethanol vs SC control; p = 0.047 for total and p = 0.046 for mitochondrial MATα1 siPin1 ethanol vs SC ethanol). SC is shown in blue and siPin1 in red. i ATP levels in AML-12 cells after Pin1 silencing (n = 3 independent experiments, p = 0.034). j Triglycerides levels (n = 3 independent experiments, p = 0.004 SC ethanol vs control; p = 0.048 siPin1 control vs SC control; and p = 0.049 siPin1 ethanol vs SC ethanol) and k Oil red O staining in AML-12 after Pin1 silencing and ethanol treatment. SC is shown in blue and siPin1 in red. l Western blot of total and mitochondrial MATα1 in HepG2 cells after PIN1 WT or R68A/R69A catalytic mutant overexpression. *p < 0.05. Statistical significance was determined by using two-tailed, one-sample t-test for treatment comparisons and ANOVA test for group comparisons. Results are shown as mean ± SEM. Source data are provided as a Source data file. DDK DYKDDDDK-Tag, EV empty vector, Mut mutant, OE overexpression, SC scramble, si silencing.