Fig 1: Knockdown of SND1 or YWHAB reversed the effects of SMARCA5 siRNA on the proliferation, invasion, and apoptosis of cervical cancer cells. The HeLa cells were transfected with SMARCA5 siRNA alone or together with SND1 siRNA or YWHAB siRNA. (A-B) The expression of SMARCA5 was detected by RT-qPCR. The expression of SND1 and YWHAB was detected by western blotting, and the quantification was performed using Image J software. n = 3. *P < 0.05, **P < 0.01, one-way ANOVA. (C-E) The changes in cell proliferation, invasion, and apoptosis were detected by Cell Counting Kit-8, Transwell, and Annexin V-FITC/PI detection kit, respectively. n = 3. **P < 0.01, one-way ANOVA. (F) The expression of apoptosis-related proteins was detected by western blotting, and the quantification was performed using Image J software. n = 3. *P < 0.05, one-way ANOVA.ANOVA: analysis of variance; FITC: fluorescein isothiocyanate; NC: negative control; OD: optical density; PI: propidium iodide.
Fig 2: The expression of YWHAB was upregulated in cervical cancer cells, and SND1 upregulated the expression of YWHAB. (A) The interaction between SND1 and YWHAB was verified by co-immunoprecipitation assay. (B) The expression of YWHAB in normal cervical cell line and different cervical cancer cell lines was detected by western blotting, and the quantification was performed using Image J software. n = 3. *P < 0.05, **P < 0.01, one-way ANOVA. (C) SND1 was overexpressed or silenced in HeLa cells. The expression of YWHAB was detected by western blotting, and the quantification was performed using Image J software. n = 3. *P < 0.05, **P < 0.01, one-way ANOVA.ANOVA: analysis of variance; IgG: immunoglobulin G; NC: negative control.
Fig 3: Overexpression of SND1 promoted proliferation and invasion, and inhibited apoptosis in cervical cancer cells. SND1 was overexpressed or silenced in HeLa cells. (A) The transfection efficiency was detected by western blotting, and the quantification was performed using Image J software. n = 3. *P < 0.05, **P < 0.01, one-way ANOVA. (B-D) The cell proliferation, invasion, and apoptosis were evaluated by Cell Counting Kit 8, Transwell, and Annexin V-FITC/PI assay, respectively. n = 3. *P < 0.05, **P < 0.01, one-way ANOVA. (E) The expression of anti-apoptotic proteins was detected by western blotting, and the quantification was performed using Image J software. n = 3. *P < 0.05, one-way ANOVA.ANOVA: analysis of variance; FITC: fluorescein isothiocyanate; NC: negative control; PI: propidium iodide.
Fig 4: Overexpression of SMARCA5 inhibited cervical cancer metastasis in vivo. HeLa cells transfected with the pLO-ciR empty vector or pLO-ciR-SMARCA5 were inoculated into nude mice to establish subcutaneous xenograft tumor models. (A) The tumor volume was monitored every week. n = 3. **P < 0.01, one-way ANOVA. After the mice were sacrificed, (B) the tumor mass was measured, (C-E) the expression of SMARCA5 was detected by RT-qPCR, and the expressions of SND1, YWHAB, and anti-apoptotic proteins were detected by western blotting, the quantification was performed using Image J software. n = 3. *P < 0.05, **P < 0.01, one-way ANOVA.ANOVA: analysis of variance.
Fig 5: SND1 functioned as an RBP for SMARCA5, and the expression of SND1 was upregulated in cervical cancer cells. (A) The binding site of SMARCA5 and SND1 was predicted by starBase. (B) The expression of SND1 in normal cervical cell line and different cervical cancer cell lines was detected by western blotting, and the quantification was performed using Image J software. n = 3. *P < 0.05, **P < 0.01, one-way ANOVA. (C) RNA pull-down assay was performed to further verify the combination of SMARCA5 and SND1. n = 3. **P < 0.01, one-way ANOVA.ANOVA: analysis of variance; NC: negative control.
Supplier Page from Abcam for Anti-SND1 antibody