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. Author manuscript; available in PMC: 2022 Mar 1.
Published in final edited form as: FEBS J. 2020 Sep 25;288(6):1871–1886. doi: 10.1111/febs.15541

Figure 3. Macrophage cytokines C5a, CCL2 and CXCL1 upregulate cell proliferation of PIN cells.

Figure 3.

(A-C) Murine PIN Pr111 cells grown on matrigel in 3D were treated with the cytokines identified from Raw264.7-coculture media including C5a (A), CCL2 (B) and CXCL1 (C) at the indicated concentrations. 72 h post-treatment, the cells were fixed, permeabilized and immunostained with cyclin D1 (green). The cell nuclei were visualized by DAPI staining (blue). Cell proliferation index of Pr111 cells under these treatments was quantified. Scale bar: 25 μm, *: p<0.05 as compared to control (student t test for C5a (A); one-way ANOVA and multiple comparisons for CXCL1 (B) and CCL2 (C)), n=5. (D) Similar to A, Pr111 cells were treated with either the identified cytokines alone or any combination of the identified cytokines as indicated. Cell proliferation index under these conditions was quantified. The data was analyzed by one-way ANOVA with Tukey-Kramer post-hoc test. *: p<0.05 as compared to the control/vehicle, n=3. (E) Normal prostate and prostate cancer sample containing PIN of human tissues were immunostained for CD88 expression. The figure shown is representative of three human tissue samples. Scale bar: 50 μm.