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. 2016 Sep 7;113(38):E5618–E5627. doi: 10.1073/pnas.1608384113

Fig. 1.

Fig. 1.

Knockdown of PDGF-BB expression and inhibition of PDGFR signaling decrease stromal fibroblasts. (A) Expression levels of PDGF-BB protein in RCC-CAKI-1, NB-IMR32, and SC-A431 tumor cells in conditioned medium. (B) Tumor microvessels and stromal components. CD31+ (red) endothelial cell and NG2+ pericyte signals (green) in various tumors. Arrowheads point to vessel-associated pericytes; arrows indicate vessel-disassociated pericytes. Quantification of NG2+ pericyte area (n = 7 random fields per group). (Scale bar, 50 μm.) (C, Top) H&E staining of tumor tissues. Arrows point to tumor stroma. (Scale bar, 100 μm.) (Middle) Endomucin+ tumor vessels (red) and αSMA+ cells (green). Arrowheads point to αSMA+ myofibroblasts. (Scale bar, 100 μm.) (Bottom) Endomucin+ tumor vessels (red) and FSP1+ stromal fibroblasts (green). Arrowheads point to stromal fibroblasts. n = 4 animals per group. (Scale bar, 100 μm.) (D) Quantification of the ratio of αSMA+ signals vs. microvascular areas (n = 12 random fields per group) and the ratio of FSP1+ signals vs. microvascular areas (n = 12 random fields per group) in various human tumors. (E) CD31+ (red) endothelial cell and NG2+ pericyte signals (green), endomucin+ tumor vessels (red) and αSMA+ cells (green), and CD31+ tumor vessels (red) and FSP1+ stromal fibroblasts (green) in scrambled-shRNA and Pdgfb-shRNA–transfected SC-A431tumors. (Scale bars: left panels, 50 μm; middle and right panels, 100 μm.) (F) Quantification of microvessel density (n = 7 random fields per group), pericyte coverage (n = 7 random fields per group), αSMA+ myofibroblast signals (n = 12 random fields per group), and FSP1+ signals (n = 12 random fields per group) in scrambled-shRNA and Pdgfb-shRNA–transfected SC-A431tumors. (G) Endomucin+ tumor vessels (red) and αSMA+ cells (green) and CD31+ tumor vessels (red) and FSP1+ stromal fibroblasts (green) in imatinib- and buffer-treated SC-A431tumors. (Scale bar, 100 μm.) (H) Quantification of αSMA+ myofibroblast signals (n = 12 fields per group) and FSP1+ signals (n = 12 random fields per group) in imatinib- and buffer-treated SC-A431tumors. Data are represented as mean ± SEM.