Figure 2

Gene silencing of SPAG9 using plasmid-mediated siRNA approach. SPAG9 specific siRNA (SPAG9 siRNA) and control siRNA (scrambled SPAG9) were used to transfect MDA-MB-231 breast cancer cells (a) No reduction in SPAG9 protein was observed in cells transfected with control siRNA. However, cells transfected with SPAG9 siRNA revealed ablation of SPAG9 protein. β-Actin protein was used as a loading control. (b) Knockdown of SPAG9 inhibits cellular growth of breast cancer cells. Histogram plot clearly shows significant growth reduction (P < 0.01) in cells transfected with SPAG9 siRNA as compared to cells transfected with control siRNA. Results are representative of three independent experiments performed in triplicates. (c) SPAG9 knockdown reduces colony forming ability of breast cancer cells. MDA-MB-231 cells transfected with SPAG9 siRNA showed significantly reduced (P < 0.001) colony forming ability in 400–1200 cells in each histogram as compared to cells transfected with control siRNA. (d) A representative photomicrograph of colony forming ability treated with control siRNA or SPAG9 siRNA in 400, 800 and 1200 MDA-MB-213 cancer cells. Columns indicate mean (n = 3); bars, standard error. *; p < 0.01, **; p < 0.001 statistically significant compared with control siRNA. These results are representative of three independent experiments performed in triplicates.