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  • br Liao XZ Tao LT Liu JH Gu YY Xie


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    Figure Legends
    Figure 1 CIZ1 is up-regulated in bladder cancer specimens and cells.
    (A) CIZ1 expression in bladder cancer (n=20) and normal tissues (n=6) was determined by immunohistochemistry assays.
    (B) Quantification of CIZ1 expression in A.
    (A and B) T24 cells expressing shCtrl or shCIZ1 lentivirus were subjected to qRT-PCR and western blot analysis of CIZ1. GAPDH serves as internal control. **p < 0.01.
    (C and D) 5637 cells expressing shCtrl or shCIZ1 lentivirus were subjected to qRT-PCR and western blot analysis of CIZ1. GAPDH serves as internal control. **p < 0.01.
    Figure 3 CIZ1 knockdown suppresses the proliferation of T24 and 5637 cells.
    (A and B) Multiparametric high-content screening analysis of shCtrl or shCIZ1 T24 cells from day 1 to day 5 after seeding. (A) Representative images of HCS. (B) Quantification of the HCS as shown in A.***p < 0.001.
    (F) Cell proliferation of shCtrl or shCIZ15637 cell was determined by MTT assay.***p
    Figure 4 CIZ1 silencing induces the apoptosis of T24 and 5637 cells.
    (A and B) Flow cytometry analysis of apoptosis showed that CIZ1 knockdown induced T24 cell apoptosis. (A) Flow cytometry determination of apoptosis. (B) Quantification result of apoptosis. **p<0.01.
    (C and D) Flow cytometry determination of apoptosis showed that CIZ1 knockdown promoted the apoptosis of 5637 cells. (C) Flow cytometry examination of apoptosis. (D) Quantification result of apoptosis. **p<0.01.
    Figure 5 Dys-regulated genes in T24 cells after CIZ1 knockdown.
    Abbreviations:CIZ1, CDKN1A‑ interacting zinc finger protein 1;BC, Bladder cancer; PCNA, proliferating cell nuclear antigen;CDKN1A,Cyclin Dependent Kinase Inhibitor 1A; CCNB1, Cyclin B1; Colcemid CCND1, Cyclin D1; PI3K,phosphatidylinositol 3-kinase; m-TOR, mammalian target of rapamycin; PTEN, phosphate and tension homology deleted on chromsome ten; TSC1,tuberous sclerosis-1; PDRG1, recombinant Human p53 and DNA-Damage Regulated 1; VEGFR, Vascular Endothelial Growth Factor Receptor 2;FFPE, formalin-fixed, paraffin-embedded; ATCC, American Type Culture Collection; DMEM, Dulbecco's Modified Eagle Media; PCR, polymerase chain reaction; SYBR, synergy brands;GAPDH,glyceraldehyde-3-phosphate dehydrogenase; BCA, bicinchoninic acid; SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis; PVDF, polyvinylidenefluoride membrane; GFP, green fluorescent protein; MTT, 3-(4,5-dimethyl-2-yl)-2,5-diphenyltetrazolium bromide; DMSO, dimethyl sulfoxide; OD, optical density; PI, propidium iodide; APC, allophycocyanin.