Fig. 2

The downstream target gene of SETD8 is RRAD and negatively regulates RRAD expression. A In the gene heatmap of knockdown of SETD8 in SW1990 cells, RRAD was significantly overexpressed. In the heatmap of genes overexpressing SETD8 in Mia PaCa-2 cells, RRAD was significantly decreased. These results indicated that RRAD was a downstream target gene of SETD8 and was negatively regulated by SETD8. B qPCR analysis was performed to exam RRAD mRNA level. These results confirmed that SETD8 knockdown led to upregulation of the RRAD expression level, while SETD8 overexpression led to downregulation of the RRAD expression level. This further proves that SETD8 negatively regulates RRAD expression. C Western blot was performed to exam RRAD protein level. The results showed that SETD8 knockdown resulted in upregulation of the RRAD protein level, while SETD8 overexpression led to downregulation of the RRAD protein level. This further indicates that SETD8 negatively regulates RRAD expression. D Representative images of IHC staining of SETD8 and RRAD in PDAC tumours and their correlation. P = 0.0018. It was further shown that SETD8 was negatively correlated with RRAD. *P < 0.05; **P < 0.01