Figure 5. Protein kinase D (PKD)3 promotes fasting response by targeting PKA activity.

(A) In vitro kinase assay using recombinant, PAH, PKA, and PKD3 as indicated on the figure the phosphorylation on PAH was assessed using RXX*S/T antibody. (B) WB for indicated proteins in primary hepatocytes overexpressing PKD3ca and control cells (n = 3). (C, D, E) WB for indicated proteins and corresponding quantifications on extracts isolated from liver of mice treated with CRT0066101 inhibitor for 5 d (10 mg per kg of body weight, i.p. injection) and corresponding control animals fasted for 6 h and euthanized 10 min after glucagon injection (200 µg/kg of body weight) (n = 3). (F) WB for PKD3 and loading control GAPDH on extracts isolated from primary hepatocytes transduced with increasing MOI of adenoviral particles containing shRNA targeted against PKD3. (G) WB for indicated proteins and corresponding quantification of WBs on protein extracts isolated from primary hepatocytes transduced with adenoviral particles containing shRNA against PKD3 or control shRNA and treated with CRT0066101 inhibitor or DMSO as vehicle. (H) Blood glucose levels before and after 10 min of glucagon injection at the dose of 200 μg/kg of body weight in mice treated with CRT0066101 inhibitor for 5 d (10 mg/kg of body weight). (I) Mice were fasted for 6 h before glucagon (I) Blood glucose levels in mice deficient for PKD3 in hepatocytes and corresponding control animals fasted for the indicated times (n = 6 per group). Data are presented as mean ± SEM. *P > 0.05, ***P > 0.001 (one-way ANOVA with post hoc Tukey’s test or t test for comparison of two groups).

Source data are available for this figure.