Fig. 4. USP11 alleviates erastin-mediated degradation of LSH.

A, C WB (top) and RT-qPCR (bottom) were analyzed in USP11 overexpressing stable cell line or USP11-KO cells treated with erastin for 24 h at the indicated concentrations. Values are presented as mean ± SD (n = 3). B, D WB (top) and RT-qPCR (bottom) were analyzed in USP11 overexpressing stable cell line or USP11- KO cells treated with erastin (20 µM) at the indicated time. Values are presented as mean ± SD (n = 3). E Co-IP with LSH antibody in HCT116 cells treated with DMSO or erastin (20 µM) for 24 h (top) and with USP11 antibody in HCT116 cells treated with erastin (20 µM) for 0, 4, 8, and 12 h respectively (bottom). F USP11-KO cells transfected with the indicated plasmids were incubated with DMSO or erastin (20 µM) for 24 h and MG132 (20 µM) for 6 h. Cell lysates were collected for Ni-NTA pull-down, and ubiquitination of LSH was analyzed by WB.