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Fig. 3 | BMC Molecular Biology

Fig. 3

From: RE1 silencing transcription factor (REST) negatively regulates ALL1-fused from chromosome 1q (AF1q) gene transcription

Fig. 3

a EMSA was performed with IRDye 800-labeled NRSE site of AF1q promoter. Competition assays were performed by further adding different concentration of molar excess of unlabeled competitive oligonucleotides. b The addition of anti-REST antibody further shifted DNA–protein complex band. Lower panel showed a longer running time. c The anti-REST antibody actually immunoprecipitated REST protein. Lane 1 was input. Lane 2 was immunoprecipitates by anti-REST antibody. Lane 3 was immunoprecipitates by IgG antibody. And the lower bands were antibody heavy chain. d Anti-RNA polymerase II antibody was used to immunoprecipitate the GAPDH promoter region in CHIP assay in HEK293 cells. A pair of primers targeting GAPDH was used to amplify GAPDH. Signals amplified from input were used as size markers for PCR. IgG and H2O were used as negative controls. e Anti-REST antibody was used to immunoprecipitate the cross-linked REST-DNA complex in CHIP assay in HEK293 cells. A pair of primers targeting AF1q was used to amplify AF1q-REST. Signals amplified from input were used as size markers for PCR. IgG and H2O were used as negative controls. And anti-RNA Polymerase II antibody was used as positive control. Si-REST was from cells transfected with psiREST

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