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. 2021 Jul 20;11:706337. doi: 10.3389/fonc.2021.706337

Figure 3.

Figure 3

Pol ι activates G6PD through OGT-promoted O-GlcNAcylation. (A) The relative mRNA level of POLI and OGT in Pol ι differentially expressed TE-1 and KYSE-150 cells. (B) the protein level of Pol ι, OGT and G6PD. (C) O-GlcNAcylation of G6PD was detected after G6PD immunoprecipitation in ESCC cells. (D) the promoter of OGT (-2000 to +500 bp) was cloned into pGL-4 vector. The pGL4-OGT and internal control reporter vector pRL-TK were co-transfected into TE-1 and KYSE-150 cells. The relative OGT promoter activity was detected by dual-luciferase reporter assay. (E) Immunohistochemical staining of Pol ι, OGT and protein O-GlcNAcylation in paraffin-embedded ESCC tissues. Scale bar = 100 μm. (F) level of OGT and protein O-GlcNAcylation in ESCC and adjacent tissues based on IHC score. (G) The correlation between Pol ι and OGT, and the correlation between OGT and protein O-GlcNAcylation were evaluated based on IHC score in 114 tumor tissue samples. (H) Survival analysis based on the IHC score of protein O-GlcNAcylation in 114 ESCC samples. Kaplan–Meier survival analysis was applied. *P < 0.05, **P < 0.01, ***P < 0.001.