c met sirna  (Exosome Diagnostics)

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

Journal: Frontiers in Immunology

Article Title: Construction of disulfidptosis-based immune response prediction model with artificial intelligence and validation of the pivotal grouping oncogene c-MET in regulating T cell exhaustion

doi: 10.3389/fimmu.2024.1258475

Figure Lengend Snippet: C-MET was a tumor driver gene and could inhibit the JAK3-STAT3 pathway. (A) The live and dead cell staining by Calcein and PI, in which siRNA-c-MET treatment increases the dead cell proportion induced by cabozantinib treatment. (B) The Edu and DAPI staining of the ln299 cell line. (C) The protein expression alteration after c-MET knockdown in the ln299 cell line, in which PDL1, p-JAK3, JAK3, and pSTAT3 were down-regulated, while (D) the expression of IL2 and IFN-γ were up-regulated in the Jurkat cell line in co-culture system. *p<0.05, ***p<0.001.

Article Snippet: Ln299 cells were transfected with c-MET small interfering RNA (siRNA) (5′-AAG GAC CGG UUC AUC AAC UUC-3′) or non-targeting negative control siRNA (RiboBio, China) using LipofectamineTM 3000 (Invitrogen, USA) according to the manufacturer’s protocol.

Techniques: Staining, Expressing, Knockdown, Co-Culture Assay

Journal: Frontiers in Immunology

Article Title: Construction of disulfidptosis-based immune response prediction model with artificial intelligence and validation of the pivotal grouping oncogene c-MET in regulating T cell exhaustion

doi: 10.3389/fimmu.2024.1258475

Figure Lengend Snippet: Down-regulation of c-MET within glioma enhanced the PBMC-derived CD8+ T cell function and proportion in the co-culture system. Glioma cell line Ln299 cells were treated with c-MET siRNA for 24h and co-cultured with PBMC for another 24h. (A) WB was used to detect the relevant protein expression in Ln299 and PBMC, in which PDL1, STAT3, pSTAT3, and pSTAT3 were down-regulated in Ln299. At the same time, IL2, IFN-γ, and CXCR9 were up-regulated in PBMC. (B) ELISA was applied to detect extracellular protein levels in the co-culture system, in which IL2, IFN-γ, and CXCL9 were higher in the si-c-MET group than those in the NC group. (C) The proportion of PD1+ PBMC was decreased by the down-regulation of c-MET in ln299 a little. (D) PD1+ CD3+CD8+ T cells were reduced evidently in the si-c-MET group than those in the NC group. **p<0.01, ***p<0.001.

Article Snippet: Ln299 cells were transfected with c-MET small interfering RNA (siRNA) (5′-AAG GAC CGG UUC AUC AAC UUC-3′) or non-targeting negative control siRNA (RiboBio, China) using LipofectamineTM 3000 (Invitrogen, USA) according to the manufacturer’s protocol.

Techniques: Derivative Assay, Cell Function Assay, Co-Culture Assay, Cell Culture, Expressing, Enzyme-linked Immunosorbent Assay