Figure 2

Gene knockdown results in cell growth inhibition and suppression of DDX10 leads to increased apoptosis. (A) Realtime quantitative PCR post-transfection of esiRNAs targeting CLTC, SKA3 and DDX10 showed efficient suppression of these three genes at the mRNA level. Suppression of EPHA5 and TNIK was not able to be assessed using this technique due to poor quality of primers. (B) siRNA targeting CLTC, EPHA5, SKA3, DDX10 and TNIK transfected MCF-10A and MCF-7 cell growth in vitro relative to controls 70 h and 120 h, respectively, after transfection is reported. Data from two independent experiments are shown with error bars representing standard deviations. Apoptosis (C) and micronuclei formation (D) of cell lines after transfection with siRNA targeting DDX10 (MCF-10A, N = 2486; MCF-7, N = 2838), SKA3 (MCF-10A, N = 2823; MCF-7, N = 5035) or both siRNAs (MCF-10A, N = 2390; MCF-7, N = 4701) was shown by numbers of cell nuclei that exhibit apoptotic nuclear morphology and micronuclei, respectively, per 1000 cells. siGFP transfected cells served as controls (MCF-10A, N = 2638; MCF-7, N = 3896). Data from a representative experiment.