Supplementary Figure 1 from c-Src Modulates Estrogen-Induced Stress and Apoptosis in Estrogen-Deprived Breast Cancer Cells

PDF file - 77K, 1A, The c-Src inhibitor blocked E2 activated non-genomic pathway. MCF-7:5C cells were treated with vehicle (0.1% DMSO), E2 (10-9 mol/L), PP2 (5x10-6 mol/L), E2 (10-9 mol/L) plus PP2 (5x10-6 mol/L) respectively for 10 minutes and the cell lysates were harvested. Phosphorylated MAPK and c-Src were examined by immunoblotting with primary antibodies. Immunoblotting for total MAPK and c-Src were used for loading controls. 1B, E2 rapidly activated MAPK and c-Src. MCF-7:5C cells were treated with vehicle (0.1% EtOH) and E2 (10-9 mol/L) for different time points as indicated and the cell lysates were harvested. Phosphorylated MAPK and c-Src were examined by immunoblotting with primary antibodies. Immunoblotting for total MAPK and c-Src were used for loading controls. 1C, E2 stimulated c-Src after 24 hours treatment. MCF-7:5C cells were treated with vehicle (0.1% EtOH) and E2 (10-9 mol/L) for different time points as indicated and the cell lysates were harvested. Phosphorylated c-Src was examined by immunoblotting with primary antibody. Immunoblotting for total c-Src was used for loading control. 1D, Quantification of Annexin V binding assay. MCF-7:5C cells were treated with vehicle (0.1% DMSO), E2 (10-9 mol/L), 4-OHT (10-6 mol/L), E2 (10-9 mol/L) plus 4-OHT (10-6 mol/L), PP2 (5x10-6 mol/L), E2 (10-9 mol/L) plus PP2 (5x10-6 mol/L) respectively for 72 hours and the cells were harvested for Annexin V binding assay through flow cytometry. The percentage of Annexin V binding was compared with control. P<0.05, * compared with control. All the data shown were representative of at least three separate experiments with similar results.