Fig. 8

Troxerutin targets CD155 to mitigate OS stemness. A Illustrative workflow diagram for the screening of potential therapeutics directed against CD155. B The top ten drugs, ranked based on affinity, with lower values indicating stronger affinity. C Western blot was utilized to assess the alterations in the phosphorylation levels of SRC following the introduction of the top ten drugs, ranked by affinity, into SJSA-1 and 143B cells. D, E The CCK-8 assay is utilized to assess the resistance of OS cells to the top ten drugs ranked by affinity. F The interaction diagram between CD155 and troxerutin, depicting hydrogen bonding with solid blue lines and hydrophobic interaction with dashed gray lines. G Hydrogen bonding diagram of CD155 and SRC (overall). H–J Hydrogen bonding diagram of CD155 and SRC (details). K The co-IP experiment was conducted to investigate the interaction between CD155 and SRC in SJSA-1 and 143B cells following troxerutin treatment. L Western blot analysis was employed to assess alterations in the phosphorylated SRC, AKT, and GSK3β protein levels within OS cells following troxerutin treatment. M Western blot is utilized to assess the nuclear translocation of β-catenin in OS cells following troxerutin treatment. N qRT-PCR was utilized to assess the expression levels of stemness markers in SJSA-1 and 143B cells following troxerutin treatment. O Representative images of the in vitro sphere formation assay. P, Q The quantity of spheres in the in vitro sphere formation assay. R–T The activity of ALDH in OS cells was measured using fluorescence-activated cell sorting analysis. Representative images and quantitative data of ALDH activity results are presented. U A refined model demonstrating troxerutin’s suppressive effect on OS stemness via the CD155/SRC/β-catenin signaling axis. *p < 0.05; **p < 0.01; ***p < 0.001