Fig. 4

Western blotting analysis of SOX9 production in HEK 293 T/17 cells. The SOX9 production of cells transfected with the pcDNA3.1-NR5A1-MU decreased significantly by about 82.11% compared with that of cells transfected with pcDNA3.1-NR5A1-WT. The single pcDNA3.1-MAP3K1-MU transfection had little effect on the SOX9 production compared to pcDNA3.1-MAP3K1-WT. Compared to the single pcDNA3.1-NR5A1-WT transfection, the SOX9 protein production of cells transfected with pcDNA3.1-NR5A1-WT and pcDNA3.1-MAP3K1-WT decreased about 17.40%, which indicated that MAP3K1 can inhibit the expression of SOX9 and was consistent with our prediction. Compared to cells transfected with the pcDNA3.1-NR5A1-MU, the SOX9 protein production of cells transfected with both mutant plasmids increased 36.64%, which indicated that the effect of increasing SOX9 production became strong after MAP3K1 was mutated. EV empty vector, WT wild type, NR5A1 nuclear receptor subfamily 5 group A member 1, MAP3K1 mitogen-activated protein kinase kinase kinase 1. *P < 0.05, NS no statistical significance