Fig. 4: SSO treatment decreases proliferation and migration in RMS cell lines.

A Non-specific (NS) and SSO55 were transfected endogenously into Rh30, RD, and Rh18 cells, and RT-PCR for IR was performed after 24 h. The percent expression of IR-B is shown as PSI. The graph below the gel picture shows the quantification from n = 5, n = 3, and n = 3 experiments, respectively. p-Value Rh30 < 0.001, RD = 0.0002, Rh18 = 0.0012. B Proliferation: Rh30 cells were seeded, transfected with either NS or SSO55, and subjected to proliferation assay using the Incucyte^® software; p-value**** < 0.0001 using 2-way ANOVA. C Rh30 cells were seeded, transfected with either IR-A-GFP or control GFP and NS SSO or SSO55 and subjected to proliferation assay using the Incucyte^® software; p-value*** = 0.0005 and ** = 0.0059 using 2-way ANOVA. D Wound healing assay: Rh30 cells were seeded in a 96-well plate and transfected with either non-specific (NS) or SSO55. After 24 h, the Incucyte^® wound maker was used to make the wound and the migration of cells was measured using the Incucyte^® platform. The wound density numbers are quantified, p-value**** < 0.0001 using 2-way ANOVA. Rh30 cells were seeded in a 96-well plate and, transfected with either IR-A-GFP or GFP and NS SSO or SSO55. After 24 h, the Incucyte^® wound maker was used to make the wound and the migration of cells was measured using the Incucyte^® platform. The wound density numbers are quantified. Statistics are calculated using 2-way ANOVA, p-value**** < 0.0001. E Trans-well migration assay: Rh30 cells were transfected with NS or SSO55 and placed in the top chamber of a dual-chamber Boyden assay and exposed to an FBS gradient or not. Non-transfected Rh30 cells were used as control. Percent migration was quantified. n = 4 for each group, an asterisk indicates p = 0.05 in one-way ANOVA with Holm–Sidak post hoc analysis. Results are shown as the standard error of the mean (±SEM).