Fig. 3: K46 of Mxi1 is the major ubiquitination target residue of UBE2O. | Cell Death & Differentiation

Fig. 3: K46 of Mxi1 is the major ubiquitination target residue of UBE2O.

From: UBE2O targets Mxi1 for ubiquitination and degradation to promote lung cancer progression and radioresistance

Fig. 3

a Mass spectrometry analysis was used for identifying the ubiquitination sites in Mxi1. The di-glycine-modified K46 in Mxi1 was detected, and the corresponding peptide was LQHSKPPR. b Alignment of the K46 corresponding amino acids sequences in Mxi1 from different species. c HeLa (upper panel) or H1299 (lower panel) cells were co-transfected with vector expressing HA-UBE2O together with indicated plasmids. After 24 h, cells were lysed for immunoblot analysis (n = 3). d HeLa cells were co-transfected with the indicated plasmids for 24 h, followed by treatment with MG132 (10 μM) for 4 h prior to collection. Cells lysates were subjected to co-IP with S beads followed by western blotting (n = 3). e Left panel: HeLa cells were transfected with SFB-Mxi1-WT or SFB-Mxi1-K46R plasmid for 24 h, and then treated with cycloheximide (CHX, 20 μg/mL) for the indicated times before harvesting. Cells lysates were analyzed by immunoblotting. Right panel: Mxi1 band intensity was measured by ImageJ software (n = 3). ***P < 0.001.

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