Figure 5. Transcription factors activated in EC by high glucose form a complex with AhR.
A Nuclear extracts from glucose-stimulated HAEC were used for immunoprecipitation with anti-AhR antibody and associated active transcription factors were identified using TF/TF array (Panomics). B: Formation of a complex between AhR and Egr-1 in response to glucose was confirmed in EMSA using consensus AhR probe. One of the two complexes was prevented by the anti-Egr-1 antibody (1 μg). C, D: The complex of AP-2 and USF-2 with AhR was confirmed in EMSA using the antibodies against AhR (1 μg), AP-2 (1 μg), USF-1 (1 μg) and USF-2 (1 μg); C - consensus AhR probe, D - probe with the sequence of THBS1 promoter fragment. RL2 antibody was used in both C and D to prove that at least one of the proteins is O-glycosylated.