Figure 4.

T444 phosphorylation is essential for the regulation of histone H2B GlcNAcylation and gene transcription by AMPK. (A) OGT was knocked down in wild-type MEFs, then shRNA-resistant wild-type OGT or T444A mutant was reintroduced into these MEFs, treated with or without AICAR, and the histone H2B GlcNAcylation signal was detected by western blot. (B) OGT wild or T444A mutant MEFs were incubated with or without glucose for 48 h, and the histone H2B GlcNAcylation signal was detected as in (A). (C) The expression of histone H2B O-GlcNAcylation common target genes in Figure 4(A) was analyzed by qRT-PCR. Data analysis is explained in the Materials and Methods section. All error bars denote s.d., n = 3.