Fig. 4

Overexpression of Thbs1 inhibits insulin sensitivity in AML12 cells. A Flag-eGFP-Thbs1 plasmid and control vector were transfected into AML12 cells. The expression of eGFP-thbs1 fusion protein in AML12 was identified by western blotting and fluorescence microscopy. B Plasmid transfected AML12 cells were placed in 96-well plates, and cell viability was detected by CCK8 assay. C. Apoptosis assays of AML12 cells after plasmid transfection were performed. D Plasmid transfected AML12 cells were treated with 1 μg/ml insulin for 1 h, and the culture medium was collected for glucose uptake measurement. E–G AML12 cells were treated with 10⁸/mL sEV^AG, 10 μM LSKL and 10⁸/mL sEV^AG + 10 μM LSKL for 48 h. Cell viability, apoptosis, and insulin-induced glucose uptake were then measured. H The expression of endoplasmic reticulum stress (ERS)-related mRNA in AML12 cells after plasmid transfection was detected by RT-PCR. I The expression of ERS-related mRNA in 10⁸/mL sEV^AG and/or 10 μM LSKL treated AML12 cells was detected by RT-PCR. J The expression of ERS-related proteins in AML12 cells was detected by western blotting. Statistical data were presented as mean ± standard deviation (SD); ns, no significance; *P < 0.05, **P < 0.01, ***P < 0.001