Fig. 1

ZNF516 is a transcription repressor. a Western blotting analysis of ZNF516 protein expression. HEK293T or MCF-7 cells were transfected with empty vector or FLAG-ZNF516. Cellular proteins were extracted from indicated cell lines and western blotting was performed with a monoclonal antibody against FLAG, GAPDH, or polyclonal antibodies against ZNF516. b Subcellular localization of ZNF516 protein. The distribution of endogenous ZNF516 was detected by immunofluorescent microscopy with polyclonal antibodies against ZNF516. DAPI staining was included to visualize the cell nucleus. Scale bar 20 μm. c The schematic diagram showing the Gal4-luciferase reporters. d Transcription repression by ZNF516. HEK293T or MCF-7 cells were transfected with different amounts of Gal4-ZNF516 expression plasmids, together with the indicated Gal4-luciferase reporter. Forty-eight hours later, luciferase activity was measured. Relative luciferase activity was calculated as firefly luciferase activity divided by renilla luciferase activity and shown relative to the control (transfected with Gal4-DBD vector). The expression of Gal4-ZNF516 was shown by western blotting. e MCF-7 cells were transfected with different amounts of FLAG-ZNF516 plasmids, together with the Gal4-SV40-LUC luciferase reporter. Forty-eight hours later, luciferase activity was measured. Relative luciferase activity was calculated as firefly luciferase activity divided by renilla luciferase activity and shown relative to the control. Each bar represents the mean ± S.D. for triplicate experiments. P-values were determined by Student’s t-test. *P < 0.05