Skip to main content
. Author manuscript; available in PMC: 2019 Nov 15.
Published in final edited form as: Lab Invest. 2018 Nov 12;99(2):260–270. doi: 10.1038/s41374-018-0122-8

Figure 2. Hyperglycemic conditions increase cellular O-GlcNAcylation and promote nuclear accumulation of GLI transcription factor.

Figure 2.

A, the activity of GLI1 and GLI2 are increased in high glucose versus normal glucose conditioned cells as evidenced by their increased nuclear presence in high glucose conditioned cells. Alpha-tubulin was used as a control for the cytosolic fraction, and HDAC1 or Histone H3 was used as a control for the nuclear fraction. B, levels of GLI1, GLI2, and OGT are elevated in cells grown in high versus normal glucose while levels of OGA are decreased in those cells. C, O-GlcNAcylation is increased in cells grown in high verses normal glucose media across several cell lines. D, the levels of O-GlcNAcylated GLI1 and GLI2 are increased in cells grown in high glucose. Lysates of SUM159 cells grown in normal or high glucose conditions were immunoprecipitated (IP’d) with RL2 antibody which detects O-GlcNAc residues. The IP’d proteins were immunoblotted for GLI1 or GLI2. E, the signal for O-GlcNAcylated GLI1 is increased in cells cultured in high glucose versus normal glucose conditions. Cell lysates were immunoprecipitated for GLI1, then immunoblotted with RL2. Endogenous GLI1 was IP’d from SUM159 cells; whereas in MDA-MB-468 cells, myc-GLI1 was transfected and IP’d. Both kinds of enrichments were immunoblotted with RL2 antibody. The numbers beneath each panel in panels A, B, C represent relative signal intensity (NG versus HG) normalized to endogenous control for each panel.