double-strand sirnas for chip sr306955  (OriGene)

Journal: The Journal of Biological Chemistry

Article Title: Cabozantinib selectively induces proteasomal degradation of p53 somatic mutant Y220C and impedes tumor growth

doi: 10.1016/j.jbc.2025.108167

Figure Lengend Snippet: Cabozantinib induces CHIP-mediated degradation of p53 Y220C . A , COV362 cells were treated with 10 μM cabozantinib for 48 h after transfected with the indicated siRNA for 24 h. P53 and MDM2 were examined by immunoblotting. GAPDH was used as a loading control. B , COV362 cells were treated with 10 μM cabozantinib for 400 h after transfected with the indicated siRNA for 24 h and 10 μM MG132 was added 8 h before harvesting the cells. Cell lysates were immunoprecipitated with an anti-p53 antibody. The immunoprecipitates and input were probed for HA and p53 by immunoblotting. C , COV362 cells were treated with 10 μM cabozantinib for 48 h after transfected with the indicated siRNA for 24 h. P53 and CHIP were examined by immunoblotting. GAPDH was used as a loading control. D , COV362 cells were treated with 10 μM cabozantinib for 48 h after transfected with the indicated siRNA for 24 h and 10 μM MG132 was added 8 h before harvesting the cells. Cell lysates were immunoprecipitated with an anti-p53 antibody. The immunoprecipitates and input were probed for HA and p53 by immunoblotting. E , COV362 cells transfected with indicated siRNA for 24 h were treated with DMSO or 10 μM cabozantinib for 48 h and then treated with 50 μg/ml cycloheximide (CHX) at different time points before harvesting the cell for immunoblotting. GAPDH was used as a loading control. Bars are mean ± SD (n = 3).

Article Snippet: Double-strand siRNAs for CHIP, USP7, c-Met, VEGFR2, and MDM2 were purchased from GenePharma.

Techniques: Transfection, Western Blot, Control, Immunoprecipitation