Fig. 4

G6PD mediated PPP activation by DCBLD1. A A histogram depicting the downregulated levels of pentose phosphate pathway metabolites in HeLa cells expressing shDCBLD1 compared to shCtrl cells. B Determination of the mRNA levels of pentose phosphate pathway-related enzymes in transduced cells by qPCR. C, E Immunoblotting was used to detect DCBLD1, His, 6PGD, and G6PD levels in cell lysates. GAPDH was used as a loading control. D, F G6PD activity was examined in these cells. G IHC staining for DCBLD1 and G6PD was performed on tumor sections obtained from a xenograft tumor model. H IHC staining for DCBLD1 and G6PD was perform on tumor sections of human cervical cancer and corresponding adjacent normal tissue. I IHC staining for DCBLD1 and G6PD was performed on tumor tissue sections from the xenograft tumor model at the same site. The area of optical density/positive staining area was quantified using ImageJ and further analyzed using GraphPad Prism. J DCBLD1 and G6PD correlation analysis using TCGA database. Data are presented as mean ± SD. Statistical significance was assessed using the multiple unpaired t test (B), the unpaired t test (D and F), a one-way ANOVA with Tukey's multiple comparisons test (G), and correlation analysis (H and I). (***, p < 0.001; **, p < 0.01; NS, no significance). Scale bar: 50 µm