Figure 3.

HMGA1 is required for the expression of G6PD and TGFβ1 in NSCLC. (A) Total number of genes with significant change in gene expression (P<0.05). 439 genes were upregulated and 713 genes were downregulated in H1299 cells transfected with HMGA1 siRNAs for 48 h, compared with the control. The differentially expressed genes were shown in volcano plots. (B) Heatmap of significantly differentially expressed genes (>2 fold) in HMGA1-silenced group versus control group. The fold change (fc) was normalized using (fc-mean^row)/SD^row. (C) Protein levels of TGFβ1 and G6PD in H1299 and H226 cells transfected with HMGA1 overexpression plasmids or siRNAs for 48 h were analyzed by western blotting. (D) The mRNA levels of TGFβ1 in H1299 and H226 cells transfected as described in (C) were analyzed by qPCR. *P<0.05, **P<0.01. (E) The concentrations of TGFβ1 secreted by H1299 and H226 cells transfected as described in (C) were analyzed by enzyme-linked immunoassay. *P<0.05, **P<0.01, ***P<0.001. (F) Immunofluorescence analysis of G6PD protein in H1299 cells transfected as described in (C). (G) The mRNA levels of G6PD in H1299 and H226 cells transfected as described in (C) were analyzed by qPCR. *P<0.05, **P<0.01. (H) The enzymatic activity of G6PD was measured in H1299 and H226 cells transfected as described in (C). *P<0.05, **P<0.01, ***P<0.001. (I) NADPH level was measured in H1299 and H226 cells transfected as described in (C). *P<0.05, **P<0.01.