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. 2020 Feb 28;11:1141. doi: 10.1038/s41467-020-14870-4

Fig. 3. CHIP is the E3 ligase for the ubiquitination and degradation of CBX4.

Fig. 3

a Lysates from HEK293T cells were IP with anti-Flag agarose and subjected to SDS-PAGE and Coomassie staining. The indicated bands were sequenced by mass spectrometry (MS) analysis. b–d HEK293T cells transfected with the indicated plasmids or siRNAs for 48 h were analyzed Western blotting. These resluts are repeated of three independent experiments. e, h, k HEK293T cells co-transfected with the indicated plasmids for 36 h were incubated with 20 μg/ml cycloheximide (CHX) for the indicated periods and then analyzed by Western blotting. Quantitation of Flag-CBX4 protein levels was based on the Western blotting results. n = 1. f, g, j HEK293T cells were co-transfected with the indicated plasmids for 48 h and then subjected to IP using anti-FLAG antibody followed by Western blotting. i U2OS/MTX300 cells were subjected to IP using anti-CBX4 antibody or anti-IgG antibody followed by Western blotting as indicated. These resluts are repeated of three independent experiments. Source data are provided as a Source Data file.