Fig. 4
NUR77 was required for CAMK4-mediated activation of autophagy and insulin signaling in HTR-8/SVneo cells. HTR-8/SVneo cells were transfected with CAMK4 overexpression plasmid and siNUR77 or siNC for 24 h, followed by incubation with 10–6 mol/L insulin for 48 h. A and B The mRNA and protein levels of NUR77 in HTR-8/SVneo cells were detected by real-time PCR (A) and western blot (B), respectively. C The proliferation of HTR-8/SVneo cells was determined by CCK-8 assay. D The invasive capacity of HTR-8/SVneo cells was determined by matrigel-based transwell assay. Scale bar = 100 μm. E The expression of autophagy-associated proteins in HTR-8/SVneo cells was detected by western blot. F Glucose uptake ability was assessed in cells of each group. G–J The ratio of Phospho.IRS-1Ser307/Total.IRS-1 (G), Phospho.IRS-1Tyr612/Total.IRS-1 (H) or Phospho.AKTSer473/Total.AKT (I) in HTR-8/SVneo cells was detected by western blot. Representative western blot images of phosphorylated IRS-1 at Ser307, phosphorylated IRS-1 at Tyr612, total IRS-1, phosphorylated AKT at Ser473, total AKT were displayed (J). Error bars depict the standard deviation of the mean. IR, insulin resistance; −, no treatment; *p < 0.05,**p < 0.01, ***p < 0.001