Figure 7
Effect of HIF-1α siRNA on mRNA expression in LTLTCa cells. A) LTLTCa cells were plated in passage media and then treated with two siRNAs for HIF-1α for 48 hours. Total mRNA was extracted and HIF-1α and BCRP mRNA, and 18S rRNA were analyzed by real-time RT-PCR. Real-time results are expressed as the mean fold-change in mRNA levels compared with negative control after normalization to 18S rRNA (mean ± SD, n = 6 independent samples/group; *versus negative control, P = 0.0057 for HIF-1α; P = 0.0026 for BCRP; one-way ANOVA). B) LTLTCa cells were plated in passage media and then treated with two siRNAs for HIF-1α for 48 hours. Total protein was extracted and HIF-1α and β-actin protein were analyzed by Western blot. Shown are representative blots and overall densitometry results of n = 6 independent cell samples/group. Densitometry results are expressed as mean fold-change in protein levels compared to negative control after normalization to β-actin (mean ± SD, n = 6 independent cell samples/group; *versus negative control (NC), P <0.0001, one-way ANOVA). ANOVA, analysis of variance; BCRP, breast cancer resistant protein; HIF-1α, hypoxia inducible factor 1 α subunit; n, number; SD, standard deviation.