Fig. 5

CAMK4 overexpression promoted invasion and autophage by upregulating NUR77 expression in primary trophoblast cells under IR conditions. A The mouse primary trophoblast cells were identified by staining of CK7. The trophoblast cells were infected with lentivirus vector carrying CAMK4 encoding sequence or/and knockdown sequence targeting mus NUR77, followed by incubation with or without 10^–6 mol/L insulin for 48 h. B and C The mRNA and protein levels of NUR77 were determined in trophoblast cells with IR treatment and silencing of NUR77. D and E The mRNA and protein levels of CAMK4 in trophoblast cells with IR treatment and CAMK4 overexpression. F and G The mRNA and protein levels of NUR77 were assessed by real-time PCR (F) and western blot (G), respectively. H The proliferation of trophoblast cells was measured by CCK-8 assay. I and J The invasive capacity of trophoblast cells was examined by matrigel-based transwell assay. Scale bar = 100 μm. K Representative western blot images and relative protein levels of Beclin 1, LC3 I/II in trophoblast cells were exhibited. L Glucose uptake ability was assessed in trophoblast cells with CAMK4 overexpression, NUR77 knockdwon or/and IR induction and insulin stimulation. Error bars depict the standard deviation of the mean. IR, insulin resistance; −, no treatment; *p < 0.05,**p < 0.01, ***p < 0.001