Fig 2: Co-inhibition of c-MET (SU11274) and PARP (NU1025) sensitizes GC cells after knockdown BRCA1/2. Knocking down BRCA1 or BRCA2 sensitizes cells to PARP and c-MET inhibition in HS746T cells expressing low levels of c-MET (AGS cells, c-MET knockdown Hs746T cells) to PARP inhibition. A, HS746T cells, control-siRNA-Hs746T cells and siBRCA1/2-Hs746T (upper panel) and AGS (lower panel) cells were exposed to 5 µmol/L of NU1025 and/or 5 µmol/L of SU11274 for 48 h for determination of cell viability (MTT metabolic activity assay). Results are expressed as percentages. Average values of three experiments ± SD are shown; (B) Western blot analysis of PARP and cl.caspase-3 in Hs746T-control-siRNA, siBRCA1/2-Hs746T (upper panel) and AGS (lower panel) cell lines. Cells were cultured with the indicated drugs (5 µmol/L NU1025, 5 µmol/L SU11274 alone or in combination for 24 h of treatment). Protein levels were normalized against actin

Fig 3: Low levels of c-MET partially sensitize GC cell lines in PARP inhibition. A, HS746T/AGS cells, control-siRNA-Hs746T/AGS cells and si-c-MET Hs746T/AGS cells were exposed to increasing doses (0-40 µmol/L) of NU1025 for 48 h for determination of cell viability (MTT metabolic activity assay). The protein levels of c-MET expression (by Western blot analysis) revealed down-regulation of the c-MET receptor in both cell lines (HS746T and AGS); (B) HS746T cells, control-siRNA-Hs746T cells and si-BRCA1/2 Hs746T cells were exposed to increasing doses (0-40 µmol/L) of NU1025 for 48 h for determination of cell viability (MTT metabolic activity assay). The protein levels of BRCA1 and BRCA2 expression (by Western blot analysis) revealed down-regulation of the BRCA1/2 in HS746T cell line; (C) HS746T cells, control-siRNA-Hs746T, siBRCA1/2-Hs746T and siMET/BRCA1/2-Hs746T cells were cultured with the indicated concentrations of NU1025 (5, 10 and 20 µmol/L) for 48 h for determination of cell viability (MTT metabolic activity assay). Error bars represent SD