Fig. 3

CD155 facilitates OS cell stemness. A Representative images depicting CSCs isolated from SJSA-1 and 143B cells. B, C The activity of ALDH in CSCs was measured using fluorescence-activated cell sorting analysis. Representative images and quantitative data of ALDH activity results are presented. D, E Utilize the qRT-PCR experiment to assess the expression of stemness markers and CD155 in CSCs cells. F Determine the protein expression level of CD155 in CSCs by western blotting. G Representative images of in vitro sphere formation assay conducted on control cells and CD155-knockdown OS cells. H The quantity of sphere in the in vitro sphere formation assay conducted on CD155-knockdown OS cells and their respective control cells. I, J The activity of ALDH in CD155-knockdown SJSA-1 and 143B cells was measured using fluorescence-activated cell sorting analysis. Representative images and quantitative data of ALDH activity results are presented. K, L qRT-PCR was utilized to assess the expression levels of stemness markers in CD155-knockdown SJSA-1 and 143B cells. M, N Utilize qRT-PCR to assess the mRNA expression levels of target genes involved in the Wnt pathway (PPARD, AXIN2, and MMP7), Notch pathway (HES1 and CCND3) and Hedgehog pathway (GLI1 and HHIP) in both control cells and CD155 knockdown OS cells. O Detection of alterations in β-catenin protein levels in OS cells following CD155 knockdown by western blotting. P Detection of alterations in β-catenin nuclear protein levels in OS cells following CD155 knockdown via western blotting. *p < 0.05; **p < 0.01; ***p < 0.001