Fig. 3: MCF2L CRE SNV1 decreases the binding affinity between MCF2L CRE and STAT3 which acts as transcriptional activator of MCF2L gene.
From: Regulatory non-coding somatic mutations as drivers of neuroblastoma
Correlation between STAT3 and MCF2L expression in a SK-N-BE(2) and b CHP-212 cells transfected with three pooled STAT3 siRNA. MCF2L and STAT3 expression relative to their respective siScramble (reported as a single scramble bar in histogram), as measured by western blot (left) and qPCR (right) 24 h, 48 h and 72 h post STAT3 silencing. β-Actin protein levels are used as loading control. c Luciferase reporter gene assay for MCF2L CRE SNV1 carried out in SK-N-BE(2) and CHP-212 72 h post STAT3 silencing. Luciferase activity of MCF2L CRE SNV1 is normalized to that from cells transfected with wild-type construct and siScramble (scramble). d Allele-specific ChIP-qPCR conducted on plasmids carrying the wild-type or MCF2L CRE SNV1 sequence upon transient transfection in SK-N-BE(2) and CHP-212 cell lines. Data is presented as fold-change of variant sequence upon comparing to wild-type sequence. All data shown are the mean ± standard deviation from three independent experiments, each done in triplicate. Significant p-values obtained by two-tailed T-test are reported by * *(* < 0.05; ** < 0.01; *** < 0.001). Ns non-significant p-value, Ref Reference, Alt Altered.
