Fig. 3.

Role of C3a in PCa progression. (A) PCa cell lines incubated in the absence and presence of C3a receptor agonist peptide (48 h) was Western blotted for cell survival and proliferation protein expression. (B) C57BL/6 mice were allografted with tissue recombinants of luciferase-expressing TRAMPC2 with wild-type (wt) or Tlr9^−/− fibroblasts. The mice were treated with saline or the TLR9 antagonist SB290157. Luciferase bioluminescence was used to image tumor progression. (C) Mean tumor volume (mm³) and SD for each treatment condition are depicted (*P < 0.05, n = 8). (D) Immunohistochemistry for phosphorylated AKT, phosphorylated histone-H3, and TUNEL staining of the tumor tissues were performed and quantitated. Hematoxylin was used as a nuclear counterstain. The corresponding graphs illustrate the mean and SD expression of the staining (n = 4). Statistical significance was determined by one-way ANOVA (*P < 0.05, **P < 0.01). (E) FACS analysis of the tumor-draining lymph nodes demonstrated that control, C3a antagonist, and TLR9-knockout fibroblasts had similar CD3⁺ T cell infiltration; however, their activation state as determined by CD8⁺/CD69⁺ expression differed significantly.