Fig. 3

The expression levels of the miR-17/92 cluster and its target genes, HSPA1B and P21, in cells after incubation with various concentrations of glucose. a Cells were cultured with 200, 900, and 1800 mg/L of glucose for 1 week and the expression of the miR-17/92 cluster (miR17-5p, miR-18a-5p, miR-19a-5p, and miR-20a-5p) was analyzed. The expression levels of the target genes of the miR-17/92 cluster (HSPA8 and P21) were examined and the expression levels of mRNA (qPCR) and protein (western blotting) of HSPA8 (b) and P21 (c) were analyzed. The graph shown under the western blotting images indicates the band densitometry results. The cell cycle was analyzed with a flow cytometer after the cells were stained with propidium iodide (d). Error bars indicate the standard error (SE) of four different experiments. In the case of the western blotting, the SE was calculated from the densitometry data. *p < 0.05, **p < 0.01, ***p < 0.001 (n = 4 each)