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. 2012 Jan 3;7(1):e29807. doi: 10.1371/journal.pone.0029807

Figure 3. Effects of HIF1α and HIF2α knockdown on the stimulation of HIF-target gene expression by CoCl2.

Figure 3

INS-1E cells (70% confluence) were transfected for 24 h with siRNA duplexes directed against Luciferase, Hif1α and Hif2α mRNA (siLuc, siHif1α and siHif2α). Then, the transfection medium was replaced with RPMI medium containing 10% foetal calf serum and G2 alone or with 200 µmol/l CoCl2. After 18 h culture, the medium was collected for insulin concentration determination and cells were processed for measurement of gene mRNA levels. Gene to Tbp mRNA ratios were expressed relative to the ratio in INS1-E cells treated with siLuc and cultured in G2. Data are means ± SEM for 3 experiments. *, p<0.05 for the effect of CoCl2 and #, p<0.05 for the effect of siRNA treatment (two-way ANOVA+test of Bonferroni).