Fig. 2: CARM1 is a binding partner of TRIM47. | Cell Death Discovery

Fig. 2: CARM1 is a binding partner of TRIM47.

From: Arginine methylation-dependent TRIM47 stability mediated by CARM1 promotes the metastasis of hepatocellular carcinoma

Fig. 2

A HEK293T cells were transfected with FLAG-TRIM47 for 48 h. Cell extracts were subjected to immunoprecipitated with anti-FLAG antibody or a control IgG. CARM1 was identified via mass spectrometry. B HEK293T cells were co-transfected with FLAG-TRIM47 and MYC-CARM1 for 48 h. Total cell extracts were immunoprecipitated with anti-FLAG or anti-MYC antibodies. FLAG-TRIM47 and MYC-CARM1 were detected by western blot. C Western blot analysis of TRIM47 protein levels in CARM1 stably knockdown SMMC7721 cells. D Western blot analysis of TRIM47 expression in SMMC7721 cells treated with CARM1 inhibitor HY12759 for the indicated times. E Cycloheximide chase analysis of TRIM47 degradation in CARM1 stably knockdown SMMC7721 cells. The relative band intensity of TRIM47 was quantified and plotted. F Western blot analysis of TRIM47 protein levels in CARM1 stably knockdown (shCARM1#1 and shCARM1#2) or control (shNC) SMMC7721 cells treated with DMSO or 10 μM MG132 for 4 h. G CARM1 stably knockdown (shCARM1#1) or control (shNC) SMMC7721 cells were co-transfected with FLAG-TRIM47 and HA-ubiquitin plasmids for 48 h. Total cell extracts were immunoprecipitated with anti-FLAG antibody. The immunoprecipitates were detected with anti-ubiquitin and anti-FLAG antibodies. H SMMC7721 cells were transfected with FLAG-TRIM47 plasmid for 48 h and treated with HY12759 for another 8 h. Total cell extracts were immunoprecipitated with anti-FLAG antibody. The immunoprecipitates were detected with anti-ubiquitin and anti-FLAG antibodies.

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