Additional file 6 of Dysregulation and prometastatic function of glycosyltransferase C1GALT1 modulated by cHP1BP3/ miR-1-3p axis in bladder cancer
Creators: name: Tan, Zengqi
name: Jiang, Yazhuo
name: Liang, Liang
name: Wu, Jinpeng
name: Cao, Lin
name: Zhou, Xiaoman
name: Song, Zhihui
name: Ye, Zhenyu
name: Zhao, Ziyan
name: Feng, Hui
name: Dong, Zewen
name: Lin, Shuai
name: Zhou, Zhangjian
name: Wang, Yili
name: Li, Xiang
name: Guan, Feng
Publisher: name: figshare
Year: 2022
Subjects: Biophysics
Biochemistry
Medicine
Cell Biology
Molecular Biology
Biological Sciences not elsewhere classified
Cancer

description: Additional file 6: Fig. S1. Expression of C1GALT1 in BLCA cells and tissues. A C1GALT1 mRNA expression in various stages of BLCA patients, in TCGA database. B-C Overall survival of dichotomized C1GALT1 (B) and T antigen (C) expression in BLCA patients using TMA. D C1GALT1 expression and T antigen levels in various BLCA (5637, RT4, KK47, J82, T24, YTS-1) and normal uroepithelial (HCV29, HUC-1) cell lines. Fig. S2. Effects of C1GALT1 silencing on malignant behavior of YTS-1 and T24 cells. (A) T antigen levels in C1GALT1-silenced YTS-1 cells by flow cytometry. (B) FGFR3 expression in C1GALT1-silenced YTS-1 cells. (C) Doxorubicin resistance of control and C1GALT1-silenced YTS-1 cells. (D) C1GALT1 expression in C1GALT1 silenced T24 cells. (E, F) Proliferation (E) and migratory ability (F) of C1GALT1-silenced YTS-1 cells. (G) C1GALT1 expression in ITZ-treated YTS-1 cells. (H-K) Proliferation (H), colony formation (I), migratory ability (J), and doxorubicin resistance (K) of ITZ-treated YTS-1 vs. control cells. Fig. S3. Effects of C1GALT1-overexpressing on malignant behavior of HCV29 cells. (A) C1GALT1 expression in C1GALT1-overexpressing HCV29 cells. (B-D) Proliferation (B), colony formation (C) and migratory ability (D) of C1GALT1-overexpressing HCV29 cells. Fig. S4. Immunohistochemical analysis of tumors of mice model. (A) Immunohistochemical analysis of Ki67, TUNEL, C1GALT1, and T antigen in tumors of mouse injected with C1GALT1-silenced, ITZ-treated, and control YTS-1 cells. (B) Immunohistochemistry analysis of Ki67, TUNEL and C1GALT1 in PDXs with or without ITZ treatment. Fig. S5. Immunohistochemical analysis of C1GALT1, T antigen, Ki67, and TUNEL in tumors of trans-splenic metastasis model mice injected with control and C1GALT1-silenced YTS-1 cells. Fig. S6. H&amp;E staining and immunohistochemistry analysis of LNs of popliteal lymphatic metastasis mouse model. (A, B) H&amp;E staining (A) and immunohistochemistry analysis (B) of LNs of popliteal lymphatic metastasis model mouse injected with C1GALT1-silenced and control YTS-1 cells into footpads. Fig. S7. Effects of miR-1-3p overexpressing on malignant behavior of BLCA YTS-1 cells. (A) C1GALT1 expression in miR-1-3p-overexpressing T24 cells. (B, C) Proliferation (B) and migratory ability (C) of miR-1-3p-overexpressing T24 cells. (D) Doxorubicin resistance of control and miR-1-3p-overexpressing YTS-1 cells. (E) C1GALT1 and T antigen levels of YTS-1 cells transfected with miR-1-3p mimic or inhibitors. (F, G) Proliferation (F) and migratory ability (G) of YTS-1 cells transfected with miR-1-3p mimic or inhibitor, in comparison with control cells. (H) Immunohistochemical analysis of Ki67, TUNEL, C1GALT1, and T antigen in tumors of mouse injected with miR-1-3p-overexpresssing and control YTS-1 cells. Fig. S8. Immunohistochemical analysis of C1GALT1, T antigen, Ki67, and TUNEL in tumors of trans-splenic metastasis model mice injected with control and miR-1-3p-overexpressing YTS-1 cells. Fig. S9. Confirmation of circRNA backsplice junctions by Sanger sequencing. (A) Sequences of backsplice junctions of selected circRNAs. Arrows: backsplice sites. (B) Production of cHP1BP3 (schematic). Fig. S10. Expression of circular cHP1BP3 and its linear counterpart in BLCA cells. (A) Sequence of mutated cHP1BP3 binding sites targeted by miR-1-3p. (B) RNA FISH for cHP1BP3 in BLCA and adjacent normal tissues. Nuclei were stained with DAPI, and miR-1-3p was FITC-labeled. (C) Expression of linear counterpart of cHP1BP3 in BLCA patients, in TCGA database. Fig. S11. Effects of cHP1BP3 silencing on malignant behavior of HCV29 and T24 cells. (A-C) Expression of cHP1BP3 (A), C1GALT1, and T antigen (B, C) in cHP1BP3-silenced YTS-1 and T24 cells. (D) Doxorubicin resistance of control and cHP1BP3-silenced YTS-1 cells. (E, F) Proliferation (E) and migratory ability (F) of cHP1BP3-silenced T24 cells. (G) Immunohistochemical analysis of Ki67, TUNEL, C1GALT1, and T antigen in tumors of mouse injected with control and cHP1BP3-silenced YTS-1 cells. Fig. S12. Immunohistochemical analysis of C1GALT1, T antigen, Ki67, and TUNEL in tumors of trans-splenic metastasis model mice injected with control and cHP1BP3-silenced YTS-1 cells.