Fig. 2: CTTNBP2 CRE SNV2 reduces the binding affinity between CTTNBP2 CRE and SIN3A which acts as transcriptional repressor of CTTNBP2 in SH-SY5Y cells.
From: Regulatory non-coding somatic mutations as drivers of neuroblastoma
Correlation between SIN3A and CTTNBP2 expression in a LAN-2 and b SH-SY5Y transfected with three pooled SIN3A siRNA. CTTNBP2 and SIN3A expression relative to their respective siScrambles (reported as single scramble bar in histogram), as measured by western blot (left) and qPCR (right) 24 h, 48 h and 72 h post SIN3A silencing. β-Actin protein levels are used as loading control. c Luciferase reporter gene assay for CTTNBP2 CRE SNV2 carried out in LAN-2 and SH-SY5Y 72 h post SIN3A silencing. Luciferase activity of CTTNBP2 CRE SNV2 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 CTTNBP2 CRE SNV2 sequence upon transient transfection in LAN-2 and SH-SY5Y 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.
