Fig 2: Clinical characteristics and prognostic factor PEA15 in the OS autophagy-related model(A) Kaplan-Meier survival analysis of the OS prognostic factor PEA15.(B) Risk survival status plot.(C and D) Univariate and multivariate COX regression analyses of the OS autophagy-related model and various clinical features.(E) mRNA expression levels of PEA15 in the normal osteoblast cell line hFOB1.19 and four osteosarcoma cell lines.(F and G) Validation of knockdown efficiency in U2OS cells and overexpression efficiency in MG63 cells (n = 3).(H and I) CCK-8 assays of cell proliferation following PEA15 knockdown and overexpression (n = 3).(J–M) Validation of autophagy activity in human tumor tissues and corresponding statistical analysis (n = 3).(N and O) Protein expression levels of PEA15 are validated by knockdown and overexpression experiments. The data are presented as mean ± SD. ∗∗p < 0.01, ∗∗∗p < 0.001. n.s.: not significant.

Fig 3: PEA15 modulates autophagy and apoptosis(A and B) Assessment of autophagic activity (autophagosome formation) following PEA15 knockdown and overexpression (n = 3).(C and D) Apoptosis assays were carried out to evaluate the effects of PEA15 knockdown and overexpression (n = 3).(E and F) Impact of PEA15 knockdown and overexpression on the expression of autophagy-related proteins.(G and H) Changes in apoptosis-related protein levels after PEA15 knockdown and overexpression. The data are presented as mean ± SD. ∗∗p < 0.01.

Fig 4: Knockdown of PEA15 Enhances the Anticancer Efficacy of DDP(A) Colony formation assays in control, DDP-treated, Si-PEA15, and DDP/Si-PEA15 combined treatment groups.(B) Migration and invasion assay images for the four treatment groups.(C) Fluorescence imaging of autophagic activity in the four treatment groups.(D and E) Expression levels of migration/invasion-related proteins and autophagy-related proteins in the four treatment groups.(F) Knockdown of PEA15 enhances the anticancer effect of DDP through the TNF signaling pathway.(G) The protein expression of ATG5 and LC3B was assessed in U2OS cells treated with the TNF-α antagonist (R-7050).

Fig 5: In Vivo Tumor Formation Assay(A) Images of solid tumors from control, DDP-treated, Sh-PEA15, and DDP/Sh-PEA15 combined treatment groups.(B and C) Statistical analysis of tumor volume and tumor weight in the four treatment groups (n = 3).(D) Hematoxylin and eosin (H&E) staining of tumors from the four treatment groups (n = 3).(E) Expression of Ki67 in tumors from the four treatment groups in the animal experiment.(F) Expression of LC3B in tumors from the four treatment groups in the animal experiment. The data are presented as mean ± SD. ∗∗p < 0.01, ∗∗∗p < 0.001.