Fig. 5: SAPS3 regulates cellular metabolism via AMPK.

a pAMPK and downstream pACC were tested by Western blotting in WT and SAPS3 KO MEF cultured in 25 mM and 10 mM glucose medium. The results are representative of three independent experiments. b WT and SAPS3 KO MEF cell lysates were immunoprecipitated with anti-PP6C antibody followed by Western blotting. The results are representative of three independent experiments. c Glucose uptake was measured using the Nova Bioprofile 100 analyzer. Mean ± s.d., n = 4 biological replicates analyzed by two-tailed t-test, ***p = 0.0006. d U-¹³C₆ glucose- derived pyruvate and glyceraldehyde-3-phosphate (GADP) levels in WT and SAPS3 KO MEF cells. Mean ± s.d., n = 3 biological replicates analyzed by two-tailed t-test. Pyruvate, *p = 0.018; GADP, *p = 0.028. e Fatty acid oxidation rate was measured using the XF-24 Seahorse system. Mean ± s.d., n = 5 biological replicates. f U-¹³C₆ glucose-derived palmitate, stearate, palmitoleate, and oleate levels in WT and SAPS3 KO MEF cells. Mean ± s.d., n = 3 biological replicates analyzed by two-tailed t-test. Palmiate, *p = 0.049; Stearate, *p = 0.022; Palmitoleate, **p = 0.005; Oleate, **p = 0.005. g AMPKα1 and α2 expression in Crispr AMPK knockout MEF cells. Mean ± s.d., n = 3 biological replicates analyzed by one-way ANOVA, **p = 0.0015; ****p < 0.001. h U-¹³C₆ glucose-derived pyruvate and phosphoenolpyruvate (PEP) levels in WT and SAPS3 KO MEF cells with or without AMPK. Mean ± s.d., n = 3 biological replicates analyzed by two-tailed t-test. Pyruvate, **p = 0.008; ***p = 0.001; PEP, WT, *p = 0.039; KO, *p = 0.032. i U-¹³C₆ glucose derived palmitate, stearate, palmitoleate, and oleate levels in WT and SAPS3 KO MEF cells with or without AMPK. Mean ± s.d., n = 3 biological replicates analyzed by two-tailed t-test. Palmitate, *p = 0.02; Stearate, **p = 0.003; Palmitoleate, *p = 0.025, ***p = 0.001; Oleate, WT, *p = 0.025; KO, *p = 0.024.