Fig. 4: K716 of HER2 is the ubiquitination target by SMURF1.
From: JAC1 suppresses proliferation of breast cancer through the JWA/p38/SMURF1/HER2 signaling
a Data from the PhosphoSitePlus (https://www.phosphosite.org) showed the potential sites required for ubiquitination of HER2. b SKBR3 cells were transfected with His-HER2 (WT) or 11 mutants (10 separate single amino acid mutants and all 10 mutated amino acids in HER2), treated with the 10 μM JAC1 for 24 h, followed by exposure to CHX (100 μg/mL) for 9 h. Protein expression of HER2 was detected by western blot. c BT474 cells were transfected with HA-HER2 (WT), HA-HER2 (K716) or HA-HER2 (ALL) for 72 h, treated with the 10 μM JAC1 for 24 h, followed by exposure to 100 μg/mL of CHX for 9 h; the protein expression of HER2 was determined by western blot. d SKBR3 cells were co-transfected with His-Ub, HA-HER2 (WT), or HA-HER2 (K716) for 72 h, followed by pretreatment with MG132 (10 μM) for 6 h. Ubiquitinated HA-HER2 was determined by IP followed immunoblot. e Colony formation assay was completed for WT/K716R HER2 transfected BT474 cells and followed by JAC1 (10 μM) treatment. f The quantitative data of colony numbers.
