Fig. 6
NUSAP1 contributes to the activation of canonical Wnt/β-catenin signaling. a GSEA plot showing that NUSAP1 expression positively correlates with Wnt/β-catenin-activated gene signatures (LABBE_WNT3A_TARGETS_UP). b TOP/FOP luciferase assay of TCF/LEF transcriptional activity in the indicated cells. c Western blotting analysis of β-catenin expression in the cytoplasm and nucleus of the indicated cells. GADPH and p84 were used as loading controls for the cytoplasmic and nuclear fractions, respectively. d Western blotting analysis of the protein expression levels of the β-catenin target genes in the indicated cells. GADPH was used as the loading control. e Lysates from Siha cells transfected with HA-TCF4 and Flag-SUMO1 were immunoprecipitated with anti-HA affinity agarose, followed by western blot analysis of TCF4 and SUMO1. f, g Lysates of Siha cells were immunoprecipitated using NUSAP1 or RanBP2 antibodies, and the interactions of NUSAP1 with TCF4 and RanBP2 were analysed by immunoblotting. h Siha cells were transfected with the indicated control RNA and siRNA against RanBP2. Lysates were immunoprecipitated with anti-HA antibody, followed by western blot analysis of TCF4 and SUMO1. i. TOP/FOPluciferase assay of TCF/LEF transcriptional activity in the NUSAP1-ovexxpressed Siha cells transfected with the indicated control RNA and SiRNA against RanBP2.All experiments were performed in triplicate. Results are the mean ± SD. *p < 0.05; **p < 0.01; ***p < 0.001