Fig. 5
PKCβ1 is involved in the adipoR1-induced IT response. A Determination of mRNAs of classic PKC isoforms (PKCα, PKCβ1, PKCβ2 and PKCγ) in the TG of mice. No signal was detected in the reactions without RT (− RT). B Bar graph revealing the effect of 100 nM adiponectin on IT in the presence of LY333531 (200 nM, n = 10 cells), HBDDE (2 µM, n = 8 cells), PKCβ1 inhibitory peptide (PKCβ1-IP, 10 µM, n = 9 cells), or PKCβ2-IP (10 µM, n = 9 cells). ***p < 0.001 (vs. control), paired t test. C Protein abundance of PKCβ1 in TG cells treated with NC-siRNA or PKCβ1-siRNA. Representative blots of at least 3 independent experiments are shown. **p < 0.01 (vs. NC-siRNA), unpaired t test. D Bar graph revealing that treatment with PKCβ1-siRNA prevented the adiponectin-induced IT increase (n = 9 cells). Adiponectin at 100 nM still significantly enhanced IT in cells transduced with NC-siRNA (n = 12 cells). *p < 0.05 (vs. control + NC-siRNA), unpaired t test. E Immunofluorescence analysis of PKCβ1 translocation mediated by 100 nM adiponectin. Arrows in white indicate the line-scanned area. Data are representative of 3 independent experiments. F Immunoblot analysis of PKCβ1 expression in cytoplasmic and membrane fractions isolated from TG cells treated with 100 nM adiponectin. GAPDH served as a control for protein loading. α-Na+/K+ ATPase was used as an indicator for membrane contamination of cytosolic extracts. Representative blots of at least 3 independent experiments are shown. **p < 0.01 (vs. control), unpaired t test