Journal: Dose-Response
Article Title: TSN Disrupts Fanconi Anemia Pathway Activation Through JAK/STAT1-Mediated Transcriptional Repression of FA Core Subunits in Bladder Cancer
doi: 10.1177/15593258251406039
Figure Lengend Snippet: Toosendanin inhibits JAK/STAT1 pathway. (A-C) Western blot shows the phosphorylation of SMAD2, STAT1 and ERK in bladder cancer T24 (A), RT4 (B) and J82 (C) cell lines induced by UVC and treated with TSN. (D-F) Immunofluorescence shows the nuclear translocation of p-SMAD2 (D), p-STAT1 (E) and p-ERK (F) in bladder cancer T24 cells induced by UVC and treated with TSN. (G-I) Western blot shows the phosphorylation of STAT1 and the expression of FANCA, FANCC, FANCF and FANCM in bladder cancer T24 (G), RT4 (H) and J82 (I) cell lines induced by UVC and treated with TSN, JAK/STAT1 agonist or inhibitor. (J-L) Histogram shows the RNA levels of FANCA, FANCC, FANCF and FANCM in bladder cancer T24 (J), RT4 (K) and J82 (L) cell lines induced by UVC and treated with TSN as well as JAK/STAT1 agonist and inhibitor by qPCR. (M-O) ChIP-qPCR shows the binding capacity of STAT1 on promoters of FANCA, FANCC, FANCF and FANCM in bladder cancer T24 (M), RT4 (N) and J82 (O) cell lines induced by UVC and treated with TSN and JAK/STAT1 agonist. Numerical P -values indicate the statistical significance compared with cells treated with UVC and TSN. Abbreviation: JI, JAK/STAT1 inhibitor Ruxolitinib; JA, JAK/STAT1 agonist IFN-γ
Article Snippet: Membranes were blocked with 5% non-fat milk in TBST for 1 h at room temperature and incubated overnight at 4°C with primary antibodies against FANCI (20789-1-AP, Proteintech, China, 1: 2000), FANCD2 (NB100-316, Novus Biologicals, USA, 1: 2000), β-Actin (AC026, ABclonal, China, 1: 2000), FANCL (A6812, ABclonal, 1: 2000), FANCB (A2435, ABclonal, 1: 2000), FAAP100 (A8591, ABclonal, 1: 2000), FANCA (A9529, ABclonal, 1: 2000), FANCC (A1812, ABclonal, 1: 2000), FANCF (18060-1-AP, Proteintech, 1: 2000), FANCG (A6206, ABclonal, 1: 2000), FANCM (A7602, ABclonal, 1: 2000), FAAP24 (26431-1-AP, Proteintech, 1: 2000), p-SMAD2 (80427-2-RR, Proteintech, 1: 2000), SMAD2 (67343-1-Ig, Proteintech, 1: 2000), p-STAT1 (82674-10-RR, Proteintech, 1: 2000), STAT1 (66545-1-Ig, Proteintech, 1: 2000), p-ERK1/2 (80031-1-RR, Proteintech, 1: 2000), ERK1/2 (11257-1-AP, Proteintech, 1: 2000).
Techniques: Western Blot, Phospho-proteomics, Immunofluorescence, Translocation Assay, Expressing, ChIP-qPCR, Binding Assay
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