Extended Data Fig. 10. Modulation of O-GlcNAc modification level with nUbc-splitOGA on c-Fos-Ubc in comparison to OGT inhibition.
Immunoblotting analysis of protein expression and O-GlcNAcylation status of c-Fos-Ubc and endogenous c-Jun under the indicated treatments corresponding to Fig. 5d, e. by either enrichment against EPEA-tag (a) or chemoenzymatic labeling followed with Biotin-IP (b). Endogenous c-Jun shows negligible changes on O-GlcNAcylation status with the co-expression of nUbc-splitOGA but shows reduced O-GlcNAc modification upon OGT inhibition with OSMI-4b. No detectable endogenous c-Fos was observed in HEK 293T cells. The data in a and b are representative of two biological replicates.