1. BCL-3 regulates expression of stemness-associated Wnt targets.
BCL-3 regulates expression of stemness-associated Wnt targets. (A) Quantitative reverse transcriptase-PCR (qRT-PCR) mRNA analysis of Wnt target gene expression in LS174T, SW620 and SW1463 cells following BCL-3 suppression. Data shows mRNA expression of genes normalised to housekeeping genes at 72 h post-siRNA transfection, showing BCL-3-knockdown cells relative to controls. One-sample t-test. N=3, ±s.e.m. *P<0.05, **P<0.01, ***P< (B) Western analysis of LGR5 and ASCL2 expression in LS174T, SW620 and SW1463 cells following BCL-3 suppression. Expressions of total β-catenin, de-phosphorylated (active) β-catenin and cyclin D1 are also shown. α-tubulin serves as a loading control. ASCL2 expression is shown in nuclear-enriched lysates, with lamin A/C serving as a loading control. (C) LGR5 expression in LS174T, SW620 and SW1463 cells following BCL-3 suppression with two independent siRNA sequences. LGR5 and BCL-3 expression was analysed by western blot. α-tubulin serves as a loading control. (D) Western analysis of BCL-3, p52 and LGR5 expression in LS174T cells at 48, 72 and 96 h following siRNA transfection with the indicated siRNAs (50 nM). α-tubulin serves as a loading control. (E) Western analysis of LS174T cells for expression of BCL-3, p52 and LGR5 at 48, 72 and 96 h following siRNA transfection with the indicated siRNAs (50 nM). α-tubulin serves as a loading control.
Danny N. Legge et al. Dis. Model. Mech. 2019;12:dmm037697
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