Additional file 1 of Targeting Drp1 inhibits ESCC progression via the ROS-PGC1-α-Nrf1/2 pathway

Additional file 1. Table S1. Clinical characteristics of ESCC patients. Table S2. Lentivirus and sh lentivirus target sequences. Table S3. QRT-PCR primers used in this study. Table S4. Primary antibodies used for Western blotting and immunohistochemistry. Table S5. The catalogue number and company name of the reagent used in this study. Figure S1. Construction of ESCC cell lines with stable knockdown and overexpression of Drp1. A Quantitative analysis for the expression of Drp1 in paired ESCC tissues. B–E Western blottingand qRT-PCRanalysis for the expression of Drp1 in KYSE-30 and EC9706 cells with Drp1 stable overexpression and control cells. EV, empty vector; Drp1, expression vector encoding Drp1. F and G Western blottingand qRT-PCRanalysis for the expression of Drp1 in KYSE-70 cells with Drp1 stable knockdown and control cells. shCtrl, control shRNA; shDrp1, shRNA against Drp1. Data information: Graphs show mean ± SEM, two-tailed unpaired t-test. p-value from t tests. *p < 0.05; **p < 0.01; ***p < 0.001. Figure S2. Drp1 overexpression promotes metastasis of ESCC cells in vitro and in vivo. A Quantitative analysis the migration of Wound-healing assay in Fig. 2A. B Quantitative analysis the number of migrated and invaded cells of Transwell assay in Fig. 2B. C, D Quantitative analysis for the levels of EMT-related proteins in KYSE-30 and EC9706cellsin Fig. 2C. E Representative images of IHC staining of MMP2 and Drp1and levelsin paired ESCC tissues. Figure S3. Targeting Drp1 inhibits the progression of ESCC cells. A–C Quantitative analysis for the levels of EMT-related proteins in ESCC cells as indicated.. shCtrl, control shRNA; shDrp1, shRNA against Drp1; EV, empty vector; Drp1, expression vector encoding Drp1. Data information: Graphs show mean ± SEM, One - way ANOVA. p-value from t tests. *p < 0.05; **p < 0.01; ***p < 0.001. Figure S4. Drp1 accelerates ESCC cell metastasis through the ROS-PGC1-α-Nrf1/2 pathway. A–C Quantitative analysis for the levels of proteins in in Fig. 4D–F. D Wound-healing migration assay for ESCC cells treatment as indicated. Scale bars: 500 μm. E Quantitative analysis the number of migrated and invaded cells of Transwell assay in Fig. 4G. Data information: Data information: Graphs show mean ± SEM, One-way ANOVA. p-value from t tests. *p < 0.05; **p < 0.01; ***p < 0.001. Figure S5. MiR-203a-3p suppress the expression of Drp1 via targeting ROS-PGC1-α-Nrf1/2 axis. A Quantitative analysis for the levels of proteins in ESCC cells as indicated.. Data information: Graphs show mean ± SEM, One - way ANOVA. p-value from t tests. *p < 0.05; **p < 0.01; ***p < 0.001. Figure S6. MiR-203a-3p inhibits the proliferation and invasion of ESCC cells by targeting Drp1. A Quantitative analysis the number of migrated and invaded cells of Transwell assay in Fig. 6C. B Quantitative analysis the migration of Wound-healing assay in Fig. 6B. C Quantitative analysis for the levels of proteins in ESCC cells as indicated.. Data information: Graphs show mean ± SEM, One-way ANOVA. p value from t tests. *p < 0.05; **p < 0.01; ***p < 0.001.