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. 2022 Mar 6;18(5):2186–2201. doi: 10.7150/ijbs.60594

Figure 4.

Figure 4

Knockdown SENP1 inhibited CSN5 expression. A,B,C The MDA-MB-231 cells was transfected with shRNA-NC or shRNA-SENP1 by lentiviral vector. The expression of CSN5 and The mRNA levels of CSN5 was measured by qPCR. D The promotor activity of CSN5 was measured by luciferase reporter assay. E Binding site of GATA1 and C/EBPbeta with the promoter of CSN5. F, G GATA1 or C/EBPbeta interacting with the promoter region of CSN5 was determined by Chip assay. H GATA1 directly regulates CSN5 in The HEK293 cells which transfected withWT-CSN5+Vector, WT-CSN5+GATA1, MUT-CSN5+Vector or MUT-CSN5+GATA1 by Dual Luciferase Reporter Assay. I, J GATA1 were detected by western blot assay. K The cell extracts from the indicated groups were subjected to immunoprecipitation with anti-GATA1 antibody or anti-IgG antibody, followed by immunoblot with anti-SUMO1 antibody. L MDA-MB-231 cells, shRNA-NC-MDA-MB-231 cells, shRNA-SENP1-MDA-MB-231 cells were treated with N-ethylmaleimide (1 mM) for 24 h. The cell extracts from the indicated groups were subjected to immunoprecipitation with anti-GATA1 antibody or anti-IgG antibody, followed by immunoblot with anti-SUMO1 antibody. Results were mean ± SD for three individual experiments which, for each condition, were performed in triplicate. * P <0.05, ** P <0.01.