Research Paper Volume 12, Issue 17 pp 17122—17136

Figure 1. IL-6 from M2 macrophage elevates the expression of PD-L1 and miR-21 in ovarian cancer cells. (A) The markers of M2 macrophage, CD206 and CD11b, were measured by flow cytometry. (B) Protein level of PD-L1 after SKOV3 and M2 macrophage co-culture, in both cell types detected by western blotting. (C) The change of PD-L1 expression after SKOV3 co-culture with IL-6 depleted M2 macrophages was detected by western blotting. (D) The expression of PD-L1, IL-6 and PD-L1 were determined by Western blotting analysis. (E) The expression of miR-21 after SKOV3 co-cultured with IL-6 depleted M2 macrophages was determined by RT-qPCR. (F) Secretion of EVs was observed by TEM. (G) The markers for EVs isolated from SKOV3 were determined by Western blotting analysis. (H) The level of miR-21 in EVs isolated from SKOV3 co-cultured with IL-6 depleted M2 macrophages was assessed by RT- qPCR. * p < 0.05 vs. control or normal. # p < 0.05 vs. co-M2 or co-SKOV3. & p < 0.05 vs. SKOV3. Statistical comparisons were performed using unpaired t test when only two groups were compared or by Tukey’s test-corrected one-way ANOVA with when more than two groups were compared.