Fig. 6

Identification of ZMAT1 downstream effectors in Pancreatic Ductal Adenocarcinoma (PDAC). A Flow diagram of the bioinformatic analyses and selection of downstream gene candidates targeted by ZMAT1. Venn diagram depicts the overlap between 789 differentially expressed genes (DEGs) upon ZMAT1 over-expression (blue) and 1079 ZMAT1-bound genes (yellow). Among 46 overlapping genes, 35 genes were up-regulated and 11 were down-regulated in SW1990/ZMAT1-OV cells. 4 downstream candidate genes were finally selected through STRING functional enrichment analyses and correlation analyses. B RT-qPCR analyses showed the relative mRNA levels of SIRT3 in ZMAT1 over-expression and deletion cell lines. C Schematic depicting the detailed information of ZMAT1-binding sites and discovered modifs (highlighted in red) on SIRT3 gene. D ChIP-qPCR analyses of ZMAT1 binding on SIRT3 promoter in SW1990/ZMAT1-OV cells. E Luciferase activity assays was performed in 293 T cells with transient expression of the WT or MUT SIRT3 gene promoter and ZMAT1. F RT-qPCR and western blot were used to detect the deletion of SIRT3 in SW1990 cells transfected with two SIRT3-siRNA. (G-H) SW1990/ZMAT1-OV cells were transfected with SIRT3-siRNA and the mRNA and protein levels of ZMAT1, SIRT3 and p53 were analyzed by RT-qPCR (G) immunoblotting (H). I CCK-8 assays were performed on SW1990/ZMAT1-OV cells transfected with SIRT3-siRNA. K Colony formation assays were performed on SW1990/ZMAT1-OV cells transfected with SIRT3-siRNA. DEGs, differentially expressed genes. All * P-value < 0.05, ** P-value < 0.01, *** P-value < 0.001. P-values were assessed using two-tailed t-tests and ANOVA followed by Dunnett’s tests for multiple comparison in B, D, E, F, G, I and J. All figures represent mean ± SD from three independent experiments