Figure 4. Down-regulation of HEXIM1 resulted in enhanced HIF-1α–HDAC1 interaction and deacetylation of HIF-1α.
(A) Control and HEXIM1 miRNA transfected MCF7 cells were subjected to hypoxia treatment (8 h). Lysates were immunoprecipitated (IP) using anti-HIF-1α antibody and analysed by Western blotting using the indicated anti-pan-acetyl antibody. Panels are representative of two experiments. (B) MCF7 cells were subjected to hypoxia treatment (8 h). Lysates were immunoprecipitated using the indicated antibodies and analysed for co-immunoprecipitating proteins by Western blotting using the indicated antibodies. Normal rabbit immunoglobulin was used as a specificity control. Input lanes represent 25 % of the total protein. Panels are representative of three experiments. (C) In vitro translated and indicated [35S]methionine-labelled proteins were incubated with the indicated GST-fusion proteins bound to Sepharose. The Input lane represents 10 % of the total volume of in vitro translated product used in each reaction. Panels are representative of eight experiments. Molecular masses in kDa are shown next to the Western blots.