Fig. 7

Abolishing Yap1 in MΦs results in an immunosuppressive microenvironment that facilitates EOC peritoneal metastases. (A) Quantification of the CD86⁺ and CD206⁺ mean of fluorescence intensity (MFI) in the LPMs and SPMs of WT and Yap1^−/− mice (n = 4) after 28 days’ intraperitoneal (i.p.) injection of ID8 cells. (B) Distinct distributions of iNOS⁺F4/80⁺, F4/80⁺CD206⁺Arg1⁺ population densities and infiltrating CD8⁺ T cells in WT and Yap1^−/− mice after 28 days i.p. injection of ID8 cells. (C) Ki67 expression was evaluated in omental isolated from WT and Yap1^−/− mice after 28 days i.p., injection of ID8 cells. (D) Metastasis in WT and Yap1^−/− mice following i.p. injection was evaluated every 7 days by bioluminescence imaging. (E) Quantitative fluorescent analysis is calculated as the total fluorescence intensity per organ within the peritoneum post-49 days (left). The results of fluorescence quantification are presented on the right. (F) For the immunofluorescence staining, an equal volume of ascites (500 µL) from each mouse was collected and then centrifuged to harvest cell spheroids. Then, a 40 μm mesh filter was employed to recover cell clusters in reverse filtration. These cells were subsequently utilized for immunofluorescent staining of Ki67⁺F4/80^+, CD206⁺CD163⁺, CD8⁺CD163⁺, and iNOS⁺CD163⁺ cells in ascites spheroids from WT and Yap1^−/− mice (n = 4) after 49 days. Scale bar: 50 μm. (G) Quantification of the CD86⁺ and CD206⁺ MFI in peritoneal MФs (LPMs and SPMs) isolated from tumor-bearing mice (i.p. injection of ID8 cells) that were treated with vehicle or XMU MP1 (n = 4) for 28 days. (H) Distinct distributions of iNOS⁺F4/80⁺, F4/80⁺CD206⁺Arg1⁺ population densities and infiltrating CD8⁺ T cells in omental were evaluated by m-IHC after vehicle or XMU MP1 for 28-day treatment. (I) Quantification of omental metastases number (n = 4). The data were shown as the mean ± SEM and were analyzed by unpaired Student’s t-tests. *, P ≤ 0.05; **, P ≤ 0.01; ****, P ≤ 0.0001