Fig. 3

WT1-AS reciprocally control WT1 expression through a feedback loop. a Subcellular localization investigation by FISH indicated that the transcript for WT1-AS was located mainly in the nucleus of Huh7 and HepG2 cell lines, the upper panel indicated Huh7 while the down panel indicated HepG2. DAPI was used as the control for labeling the nucleus. b Sequences of the binding site for WT1-AS in the promoter region of WT1 and the WT1 promoter and a mutant sequence were cloned for further experiments. Wild type presented in blue while mutant type in red. c Bioinformatics predicted the binding site between the WT1-AS with WT1. d, e Cells were co-transfected with WT1-AS plasmid or control, Renilla luciferase vector pRL-SV40 and WT1 promoter (wild type and mutant type). Both firefly and Renilla luciferase activities were measured in the same sample. Firefly luciferase signals were normalized with Renilla luciferase signals. Cells treated with controls plasmid were normalized to 100 %. Panel d presented HepG2 cell line while panel e presented Huh7. All data presented as mean ± SD, * indicated P < 0.05 by unpaired Student’s t test