Fig. 5

PTRH1 inhibits PD-L1 expression by binding PD-L1 mRNA 3'-UTR A-C qPCR analysis A and Western blotting B and Flow cytometry analysis C of PD-L1 expression in PANC-1, SW1990, AsPC-1, and CFPAC-1 following different treatment with overexpression or knocking down of PTRH1 in 5.5 mM or 25 mM sugar concentration medium. D qPCR analysis of the PD-L1 mRNA content in RNAs obtained by pull-down of endogenous overexpressed PTRH1 protein in SW1990 cell lines. Anti-IgG antibody was used as a control. E Veen plot depicting overlapping binding peaks identified by three independent methods (Ablife, Piranha, and Cims) that PTRH1 may combine and interact to. RNA library was obtained by iRIP-seq experiment. F PTRH1 binding peaks on the PD-L1 mRNA 3′-UTR in three analytic strategies (Ablife, Piranha, and Cims). G A preferred motif that PTRH1 binding to on the PD-L1 mRNA 3′-UTR. H qPCR analysis of the binding of PTRH1 to PD-L1 mRNA based on the cDNA library obtained from iRIP-seq experiment. iRIP Improved RNA Binding Protein Immunoprecipitation, ns non-significant. The graphs show representative results from three independently repeated experiments. *p  < 0.05, **p  < 0.01, ***p  < 0.001