Fig. 6: Perturbation in phosphocreatine metabolism is linked to CCL2 production via effects on AMPK activity.
From: Impaired phosphocreatine metabolism in white adipocytes promotes inflammation
a, PRKAA1 and PRKAG1 mRNA expression in human adipocytes transfected with siPRKAA1, siPRKAG1 or siC (three replicates per condition, repeated twice). Overall P = 0.005 for PRKAA1 and P < 0.0001 for PRKAG1. b, AMPK, phosphorylated AMPK (pAMPK) and actin levels determined by western blot in human adipocytes transfected with siPRKAA1, siPRKAG1 or siC. The left panel shows representative blots and bar graphs to the right show quantifications of the indicated protein levels (three independent experiments). Overall P = 0.0013. c, Same experimental setup as in a but displaying the effects on CCL2 mRNA levels (three replicates per condition, repeated twice). Overall P = 0.0011. d, Same experimental setup as in a but displaying the effects on CCL2 secretion (three replicates per condition, repeated twice). Overall P = 0.0115. e, AMPK, pAMPK, CK-B and GAPDH determined by western blot in human adipocytes transfected with siC or siCKB. The left panel shows representative blots and bar graphs to the right show quantifications of the indicated protein levels (two replicates per condition, repeated two times). **P = 0.0018. f, AMPK, pAMPK, CK-B and Lamin A/C determined by western blot in human adipocytes transfected with siC or siCKB, after incubation for 24 h with phosphocreatine (30 mmol l−1) or PBS. The left panel shows representative blots and bar graphs to the right show quantifications of the indicated protein levels (quantifications from four independent expirements). **P = 0.0078. g, CCL2 expression in human adipocytes transfected with siC or siCKB, after incubation with vehicle (DMSO) or PF-739 (5 μmol l−1) for 24 h (three replicates per condition, repeated twice). Overall P < 0.0001. Data were analysed by Student’s two-sided t-test in e and f, by one-way ANOVA in panels a–d and two-way ANOVA in g. Tukey’s post-hoc tests indicated by *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Data are shown as mean ± s.e.m.
