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. 2021 Apr 19;7:85. doi: 10.1038/s41420-021-00426-y

Fig. 3. JAC1 downregulates HER2 expression via ubiquitin–proteasome pathway.

Fig. 3

a BT474 cells were treated with the JAC1 (0, 1, 10 μM) for 24 h, followed by Cycloheximide (CHX, 100 μg/mL) for 0, 3, 6, 9 h. HER2 protein expressions were determined by western blot. b, c Ubiquitination of HER2 was induced by JAC1. Ubiquitination of the HER2 protein was immunoprecipitated and detected a ubiquitin antibody. d The HER2-targeting E3 ubiquitin ligases were predicted using the public database website (ubibrowser.ncpsb.org/). e After BT474 cells were treated with the JAC1 (0, 1, 10 μM) for 24 h, the expressions of E3 ligases which were ranked at the top were determined by western blot. f BT474 cells were pretreated with MG132 (10 μM) for 6 h, and the endogenous protein–protein interaction between SMURF1 and HER2 was assessed by IP and followed by western blot. g Western blotting analysis of HER2 and SMURF1 in BT474 cells treated with JAC1 or si-SMURF1 and corresponding controls.