Fig. 8: In vitro validation of DCUN1D5’s tumorigenic potential.

A Growth quantification of both sensitive and drug-resistant osteosarcoma cell lines using CCK-8. *P < 0.05. B qRT-PCR assessment of DCUN1D5 levels in osteosarcoma and cisplatin-resistant cell lines. C Western blot analysis of protein expression. D Confocal imaging of DCUN1D5 in both sensitive and resistant osteosarcoma cells. E Observation of tumor spheroid formation post-DCUN1D5 knockdown in resistant cells. F 3D culture analysis post-DCUN1D5 knockdown in resistant cells. G Flow cytometric analysis of osteosarcoma cell apoptosis at 48 h. H Quantitative apoptosis assessment in osteosarcoma cells. ***P < 0.001 vs. SiRNA-NC. I a Si-NC, b Si-DCUN1D5 were co-cultured with U-2OS/CDDP cells for 24 h, c Si-NC, d Si-DCUN1D5 were co-cultured with MG63/CDDP cells for 24 h. J a Oe-NC, b Oe-DCUN1D5 were co-cultured with U-2OS/CDDP cells for 24 h, c Oe-NC, d Oe-DCUN1D5 were co-cultured with MG63/CDDP cells for 24 h. protein expression of PI3K/AKT/GSK3β signal path biomarkers were identified via immunoblotting. The quantitation data from the immunoblotting analysis were analyzed via the ImageJ program, which was expressed as mean ± SD.