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. 2023 Aug 15;19(13):4259–4277. doi: 10.7150/ijbs.85986

Figure 7.

Figure 7

PAI-1-related signalling pathways were activated after coculture. (A) Total TGFβ1 level in the culture media of monocultured tumor cells, non-activated platelets, activated platelets, and B16F10-C3 and 231-GFP cells cocultured with non-activated platelets through ELISAs. (B) Western blotting shows the levels of TGFβ/Smad pathway-related proteins. (C) The mRNA levels of PAI-1 in B16F10-C3 cells after knocking down TGFβ1, TGFBR1, TGFBR2, Smad2, Smad3 and Smad4 using siRNAs. (D) PAI-1 protein levels after knocking down targeted genes. (E) mRNA levels of PAI-1 in B16F10-C3 cells after treatment with the TGFBR1 inhibitor SB431542. (F) The protein levels of p-TGFBR1, p-Smad2/3, Smad2-4, and PAI-1 after SB431542 treatment in B16F10-C3 cells. (G) Quantified results of the Transwell migration assay in B16F10-C3 cells after SB431542 treatment. (H) The protein levels of the PI3K/AKT pathway between monocultured and cocultured B16F10-C3 cells with knockdown and overexpression of PAI-1 and recombinant TGFβ1 protein treatment. The results are displayed as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001.