Real-time RT-PCR measurements of glucose-induced changes in HIF subunits and HIF-target gene mRNA levels in INS-1E cells.

INS1-E cells were cultured 18 h in the presence of 2, 5, 10 or 30 mmol/l glucose and the mRNA levels of selected genes were measured by real time RT-PCR (insulin secretion during culture was (mean ± SE ng/h, n = 7) 24±2 in G2, 40±7 in G5, 69±15 in G10 and 57±13 in G30). Tbp mRNA levels and Gene/Tbp mRNA ratios were expressed relative to the level in G2. Data are means ± SE for 3 to 13 experiments. The cycle threshold (Ct) measured in G2 (real time PCR performed with islet cDNA equivalent to 2 ng total RNA) are shown as rough indicators of gene mRNA levels.aPbPvs. cells cultured in G2 (one-way ANOVA+test of Newman-Keuls).ND, not detected.