Fig. 7: 463aa modulated the ubiquitination modification of NR2F1. | Cell Death & Disease

Fig. 7: 463aa modulated the ubiquitination modification of NR2F1.

From: RBMS3-induced circHECTD1 encoded a novel protein to suppress the vasculogenic mimicry formation in glioblastoma multiforme

Fig. 7

A, B 463aa and NR2F1 complexes were co-immunoprecipitated with anti-463aa and anti-NR2F1 antibodies, and NR2F1 and 463aa were detected, respectively. C Relative expression of NR2F1 mRNA in 463aa overexpression GBM cells was tested by qPCR assay. D Relative expression of NR2F1 protein in U87 and U251 cells with 463aa overexpression was tested by western blot (n = 3). **P < 0.01 vs OV-NC group. E U87 and U251 cells with 463aa elevation were incubated with CHX for the indicated times. The relative expression of NR2F1 protein level was shown. F Control or 463aa-overexpressing GBM cells were treated with CHX for the indicated time, then were treated with or without the proteasome inhibitor MG132 for 2 h before collection. The expression of 463aa and NR2F1 were determined and quantified (n = 3). *P < 0.05 vs 463aa group. G HA-Ubiquitin, myc-NR2F1, or Flag-463aa plasmids were transfected alone or together into HEK293T cells as indicated. NR2F1 ubiquitination was detected by immunoprecipitation of 463aa with anti-myc antibody and western blot with anti-HA antibody. The expression of NR2F1 and 463aa in the whole-cell lysates was confirmed. H The ubiquitination of endogenous NR2F1 was analyzed by immunoprecipitation in U87 and U251 cells overexpressing 463aa.

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