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Fig. 4 | Journal for ImmunoTherapy of Cancer

Fig. 4

From: Targeting Interleukin(IL)-30/IL-27p28 signaling in cancer stem-like cells and host environment synergistically inhibits prostate cancer growth and improves survival

Fig. 4

Histopathological and cytofluorimetric aspects of the spleen of IL-30KO mice. a H&E stained sections of the spleens obtained from untreated WT and IL-30KO mice showing differences in macroscopic appearance and in size. b Total number of cells in the spleens from WT or IL-30KO mice, injected or not with 1X105 PIN-SCs. Results are reported as mean ± SD of viable cells evaluated by flow cytometry, using 7-amino-actinomycin D (7-AAD) staining. c and d illustrate flow cytometry analysis of Treg cells in one representative spleen sample from a WT mouse, injected with PIN-SCs. c. After exclusion of dead cells (7-AAD-positive), CD3 + CD4 + CD45R- nucleated cells were gated and analyzed for Foxp3. d. Most of the CD4+Foxp3+ population showed high CD25 expression. Isotype controls were used to assess the background. Experiments were performed at least in triplicate. e Flow cytometry analysis of Tregs in one representative spleen sample from an IL-30KO mouse, injected with PIN-SCs. After exclusion of dead cells (7-AAD-positive), CD3+CD4+CD45R nucleated cells were gated and analyzed for Foxp3. Experiments were performed at least in triplicate. f Immunohistochemical features of spleens obtained from healthy and (PIN-SC) tumor-bearing WT and IL-30KO mice. Results obtained from mice bearing control shPIN-SC tumors are not different from those obtained in mice bearing PIN-SC tumors. Magnification: × 400. Scale bars: 30 μm. g Immunohistochemical features of spleens obtained from WT and IL-30KO mice bearing shPIN-SC or IL-30shPIN-SC tumors. Results obtained from WT and KO mice bearing control shPIN-SC tumors are not different from those obtained in the same mouse strain bearing PIN-SC tumors. Magnification: × 400 (top); × 630 (bottom). Scale bars: 30 μm (top); 20 μm (bottom). h Double immunohistochemistry reveals that in the spleen of IL-30KO mice bearing shPIN-SC or IL-30shPIN-SC tumor, IFNγ (brown) mostly co-localizes with F4/80+ macrophages (red) and, to a lesser extent with CD3+T cells (red). Magnification: X400.Scale bars: 30 μm

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