Fig. 3
The AdipoR1-induced IT increase is mediated by the protein kinase CK2α. A Time course of IT changes (left) and bar chart (right) showing the effect of adiponectin at 100 nM on IT in cells dialyzed with GDP-β-S (1 mM) (n = 11 cells). Insets indicate representative current traces. Numbers represent points used for exemplary traces. B Coimmunoprecipitation of adipoR1 with RACK1 in the TG. Representative blots of at least 3 independent experiments are shown. C Protein abundance of RACK1 in TG cells treated with RACK1-siRNA or NC-siRNA. Representative blots of at least 3 independent experiments are shown. **p < 0.01 (vs. NC-siRNA), unpaired t test. D Summary of results revealing that treatment with either NC-siRNA (n = 10 cells) or RACK1-siRNA (n = 11 cells) did not affect the adiponectin-induced IT response. *p < 0.05 (vs. control + NC-siRNA), #p < 0.05 (vs. control + RACK1-siRNA), unpaired t test. E Interaction of adipoR1 with PKCK2α in the TG. Representative blots of at least 3 independent experiments are shown. F-G Time course of IT changes showing the effect of adiponectin at 100 nM on IT in cells preincubated with CX-4945 (F, 10 μM) or TBB (G, 20 μM). Insets indicate representative current traces. Numbers represent points used for exemplary traces. H Summary of the results revealing the effect of adiponectin at 100 nM on IT in the presence of CX-4945 (n = 8 cells) or TBB (n = 11 cells). ***p < 0.001 (vs. control), paired t test. I Protein abundance of CK2α in NC-siRNA- or CK2α-siRNA-treated groups. Representative blots of at least 3 independent experiments are shown. **p < 0.01 (vs. NC-siRNA), unpaired t test. J Representative traces (left) and bar chart (right) revealing that treatment with CK2α-siRNA prevented the adiponectin-induced IT increase (n = 11 cells). Adiponectin at 100 nM significantly enhanced IT in cells transduced with NC-siRNA (n = 12 cells). *p < 0.05 (vs. control + NC-siRNA), unpaired t test