Fig. 5

Metformin downregulated SLC7A11 expression by inhibiting UFMylation of SLC7A11. A UFM1 and SLC7A11 expression in breast cancer cell lines was detected via Western blotting. Scatter plots showing positive correlation of UFM1 and SLC7A11 expression in different cancer types. B TCGA database analyse the effect of UFM1 or SLC7A11 on survival. C After knockdown of UFM1 with siRNA, the relative SLC7A11 protein and RNA expression levels were measured via Western blotting and qRT-PCR. D The protein level of SLC7A11 in breast cancer cell lines was detected by Western blotting after cells were treated with cycloheximide (CHX, 100 μg/ml) after knockdown of UFM1. E After siRNA-mediated knockdown of UFM1, SLC7A11 UFMylation was measured via Co-IP. F SLC7A11 UFMylation was measured via Co-IP in the absence or presence of UfSP2. G SLC7A11 UFMylation was measured via Co-IP in the absence or presence of metformin