Fig. 1

REST inhibited human AF1q gene transcription. a Real time PCR showed that REST expression was decreased by psiREST. SYBR Green gene expression assay of REST was performed. 18S rRNA was used as the internal control. The values represent the mean ± SE (n = 3). *P < 0.05 by Student’s t test. Con control. b RT-PCR showed that REST expression construct pREST increased REST mRNA expression, and REST knock down construct psiREST decreased REST mRNA expression. β-actin was used as the internal control. The values represented the mean ± SE (n = 3). *P < 0.01 by Student’s t test. c The knockdown effect of psiREST was verified in protein level by Western blot. REST was detected with anti-REST antibody. β-actin was used as the internal control. The values represented the mean ± SE (n = 3). *P < 0.01 by Student’s t test. d RT-PCR showed that AF1q mRNA was repressed by REST overexpression in HEK293 cells. β-actin was used as the internal control. The values represented the mean ± SE (n = 3). *P < 0.01 by Student’s t test. e Genomic organization of AF1q gene. AF1q gene had two exons and one intron. TSS, transcription start site. f pAF1q-promoter contains the functional promoter of AF1q gene, containing the 1809-bp fragment from the human AF1q gene 5′UTR, was transfected into HEK293 cells. +1 is the first base of the first exon. Dual-luciferase activity was measured by a luminometer 24 h after transfection. The values represented the mean ± SE (n = 3). *P < 0.01 by Student’s t test. g The expression of REST was examined by Western bolt. REST was detected with anti-REST antibody. Different mass ratios were marked under the Western blot figure. β-actin was used as the internal control. h AF1q gene promoter was inhibited by REST. REST was co-transfected with pAF1q promoter into HEK293 cells. Dual-luciferase activity was measured 24 h after transfection by a luminometer. The values represented the mean ± SE (n = 3). *P < 0.01 by Student’s t test