Investigation of transcriptional repression using a panel of GAL4-PHF1b fusion proteins. A) Full length PHF1b and truncated PHF1b versions fused to GAL4 (1–100) were recruited upstream of the TK enhancer promoter which was linked to a luciferase reporter gene (pG5-200tkLUC). GAL4 (1–100) contains the DNA binding domain that recognizes GAL4 regulatory sites. GAL4-PHF1b fusions were expressed from a CMV promoter. COS-7 cells were co-transfected with the reporter plasmid (pG5-200tkLUC) and the constructs as indicated. 48 hours after transfection, cells were harvested and assayed for luciferase activity. Results shown are mean values ± SEM and normalized to protein content within each dish as well as to vector control (pG5-200tkLUC+ GAL4 (1–100) defined as 100%). “*” indicates significantly different from vector control (p < 0.05) as determined by 95% confidence interval. B) GAL4-PHF1b fusion proteins are expressed in COS-7 cells as shown by Western analysis. Extracts of COS-7 cells mock transfected or expressing PHF1b constructs were analyzed by Western analysis using a GAL4 antisera. No proteins were detected in mock-transfected cells (1) and proteins were detected for cells transfected with PHF1b (2), PHF1bΔ4 (3), PHF1bΔ3 (4), PHF1bΔ6 (5). Molecular size markers are to the right.
Saha et al. BMC Pharmacology and Toxicology 2013 14:37 doi:10.1186/2050-6511-14-37