Fig. 1

Higher NF-kB and AR expression correlates with muscle-invasive BCa in human clinical samples and ASC-J9® treatment repressed NF-κB signaling pathway in J82 and TCC-SUP cells and decreased AR expression in AR positive TCC-SUP cells. a Oncoprints of NFKB1, NFKB2, RELA, RELB, and AR genomic alterations. Individual genes are represented in each row, and individual tumors are represented as columns. Genetic alterations are color coded: dark blue, homozygous deletion; red, amplification; green, misssense mutation; black, truncating mutation; black arrowhead, protein upregulation; pink frame, mRNA upregulation; and light blue frame, mRNA downregulation. The oncoprints are based on data obtained from the cBioPortal for Cancer Genomics (http://www.cbioportal.org/). b Representative images of AR and NF-kB p65 staining in muscle invasive urothelial tumor tissues. Scale bar, 100 mm. c The Pearson correlation (shown as key) between AR and NF-kB nuclear expression. d NF-κB transactivation affected by ASC-J9® (ASC) was determined using reporter gene assay with plasmids containing luciferase gene under NF-κB response element (NRE) control in J82 and TCC-SUP cells after 24 h treatment. e The activation of NF-κB affected by ASC was measured by ELISA to determine the phospho-p65 in J82 and TCC-SUP cells after 24 h treatment. f The protein expression of IκBα affected by ASC-J9® was measured using western blotting after 48 h treatment in J82 and TCC-SUP cells. g Decreased levels of p65 protein in the nuclear fraction of J82 and TCC-SUP cells following ASC-J9® treatment. h The effect of IκB kinase (IKK) activator, prostratin, on the suppressive activity of ASC on cell growth was determined in J82 and TCC-SUP cells after 48 h treatment. i ASC-J9® decreased AR protein levels in TCC-SUP cells after 48 h treatment. Data are presented as mean ± SD; *, p < 0.05; **, p < 0.01, vs no treatment control, NS = Not Significant