Fig. 4: Eepd1 promotes thermogenesis and lipolysis of mature adipocytes through PKA signaling.
From: Myristoylated Eepd1 Enhances Lipolysis and Thermogenesis through PKA Activation to Combat Obesity
a C3H10T1/2-derived adipocytes were transfected with siRNAs and then OCR was measured by Seahorse assay (n = 6 biological replicates). b qRT‒PCR analysis of the indicated genes (n = 4 biological replicates). c C3H10T1/2-derived adipocytes were reverse transfected with mock or Eepd1 expressing plasmids and then OCR was measured by Seahorse assay (n = 7 biological replicates). d qRT‒PCR analysis of the indicated genes in C3H10T1/2-derived adipocytes transfected with plasmids (n = 6 biological replicates). e, f Glycerol and NEFA release assay in C3H10T1/2-derived adipocytes treated with NC or siEepd1 for 48 h (n = 6 biological replicates). g, h Glycerol and NEFA release assay in C3H10T1/2-derived adipocytes transfected with indicated plasmids for 48 h (n = 5 biological replicates). i Heatmaps of the leading gene expression from the pathway of regulation of thermogenesis and lipolysis in adipocytes. j KEGG and GO analysis of the data from RNA-seq. k, l Immunoblots of phospho-PKA substrates (RRXS*/T*) and HSL in iWAT and eWAT from WT and EKO mice fed a HFD for 12 weeks. β-actin was used as a loading control. m Immunoblots of phospho-PKA substrates (RRXS*/T*) and HSL in C3H10T1/2-derived adipocytes treated with NC or siEepd1 for 48 h. GAPDH was used as a loading control. n Immunoblots of phospho-PKA substrates (RRXS*/T*) and HSL in C3H10T1/2-derived adipocytes transfected with plasmids for 48 h. β-actin was used as a loading control. o Immunoblots of phospho-PKA substrates (RRXS*/T*) and HSL in C3H10T1/2-derived adipocytes incubated with H89 for 2 h and then transfected with indicated plasmids for 48 h. β-actin was used as a loading control. p OCR in C3H10T1/2-derived adipocytes incubated with H89 for 2 h and then infected with indicated adenovirus for 48 h (n = 7 biological replicates). Data presented as mean ± SEM. Significance was assessed by unpaired two-tailed Student’s t test (a–h), two-sided Hypergeometric test with Fisher’s exact test (j) or one-way ANOVA (p). OCR, oxygen consumption rate. ISO, isoproterenol. Source data are provided as a Source Data file.
