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. 2020 Feb 26;11(2):151. doi: 10.1038/s41419-020-2350-2

Fig. 6. TRIM21 is an E3 ubiquitin ligase that specifically targets SET7/9 for degradation.

Fig. 6

a MCF-7 and MDA-MB-231 cells were transfected with the vector alone or FLAG-tagged TRIM21 or were treated with control shRNA or TRIM21 shRNA. Cellular extracts were collected, and western blot was performed with specific antibodies against the indicated proteins. b The cells from the experiments in a were analyzed for the expression of SET7/9 mRNA by RT-qPCR. Error bars represent the mean ± SD of triplicate experiments. c MCF-7 cells were transfected with the vector alone or FLAG-TRIM21 and then treated with DMSO or MG132 for 6 h before the cells were collected. The cell lysates were analyzed by western blot with antibodies against the indicated proteins. d MCF-7 cells were transfected with the vector alone or FLAG-TRIM21. Forty-eight hours after transfection, the cells were treated with 50 μg/ml cycloheximide (CHX) for the indicated times, and cellular proteins were extracted for western blot analysis. e MCF-7 cells were co-transfected with the indicated plasmids. Forty-eight hours after transfection, the cells were treated with MG132 for 12 h before cellular extracts were prepared for immunoprecipitation (IP) with anti-Myc followed by immunoblotting (IB) with anti-HA.