Fig 1: 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
Fig 2: Dependence of various HIV-1 mutants on TNPO3, RanBP2 and Nup153.(A) HeLa cells were transfected with siRNA against TNPO3 and infected with VSV-G-pseudotyped GFP reporter viruses. (B) To deplete RanBP2, HeLa cells were first infected with a VSV-G-pseudotyped virus carrying a Crimson reporter and shRNA targeting RanBP2. They were then challenged with VSV-G-pseudotyped GFP reporter viruses. Viral infectivity shown in this data was determined by measuring GFP+ cells in crimson-positive populations. (C) HeLa cells were transfected with siRNA against Nup153 and infected with VSV-G-pseudotyped GFP reporter viruses. Results are one representative experiment of at least two experiments. Standard errors of a single triplicate experiment are indicated with the error bars.
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