Fig 4. MTG binds to HIF1a response elements in-vivo in hypoxia as shown by chromatin immunoprecipitation (ChIP).

ChIP assays were carried out using chromatin isolated from Raji-MTG16 cells incubated at 4% O ₂ and induced with 20 ng/ml of doxycycline 12 h to express MTG16. Schematics of promoter regions of PDK1, PFKFB3 or PFKFB4 including HRE are shown. Numbers are relative to transcription start site (TSS). Positions of primers for real time PCR are indicated. The numbers indicate nucleotide position in the promoters corresponding to TSS. The protein-DNA complexes were immunoprecipitated with control α-β-actin, α-MTG (detecting MTG16), α-HIF1α or α-HIF1β antibodies and analyzed by qPCR (mean ± SEM, n = 3). Enrichment of MTG16, HIF1α and HIF1β on HRE is shown. No enrichment was seen with control antibody or without antibody.