Fig 2: Mechanistic insights into PEA15 downstream signaling(A) Heatmap of transcriptomic analysis in U2OS cells following PEA15 knockdown.(B) CCK-8 assay was adopted to assess cell proliferation after FABP3 knockdown in U2OS cells (n = 3).(C) Colony formation assay in U2OS cells with FABP3 knockdown (n = 3).(D and E) Migration and invasion assays in U2OS cells with FABP3 knockdown, along with the expression levels of related proteins (n = 3).(F and G) Fluorescence imaging of autophagic activity and expression of autophagy-related proteins following FABP3 knockdown.(K–L) Flow cytometry analysis of apoptosis and expression of apoptosis-related markers after FABP3 knockdown (n = 3).(M) KEGG pathway analysis of transcriptomic data from U2OS cells with PEA15 knockdown.(N) In U2OS cells with PEA15 knockdown, FABP3 was overexpressed, followed by the detection of ATG5 and LC3B protein expression levels.

Fig 3: 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.

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: 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.