Fig. 7.

Effect of increasing doses of Thiamet-G (ThG) on O-linked β-N-acetylglucosamine (O-GlcNAc) modification of cytochrome c oxidase subunits. Cytochrome c oxidase complex was pulled down to assess any change in O-GlcNAcylation status in response to varying doses of Thiamet-G treatment for 6 h. As noted in materials and methods, the Millipore CTD 110.6 antibody was used in the mitochondrial O-GlcNAc blots shown here. A–F: Western blot analysis of O-GlcNAc with and without preincubation with 100 mM GlcNAc to show specificity for O-GlcNAc immunoblotted (IB) proteins (A) is represented as well as Western blot analysis of MTCO2 (B and D), MTCO1 (B and E), subunit 4-1 (B and F), and VDAC (B) including relative quantification (C) of the Western images normalized to the untreated control. IP, immmunopecipitation. Data are means ± SE; n = 3 replicates per sample. *P < 0.05, different from untreated control.