Fig. 4

Biglycan inhibition reduces stromal fibrosis and normalizes immune suppressive microenvironment. a Representative images of picrosirius red staining for collagen in E0771 tumors from WT and Bgn KO mice. Scale bar = 100 μm. b Quantification of collagen accumulation. Data represent means ± SD. n = 5 fields, 5 mice per group. c Col1a1 mRNA expression in E0771 tumor tissues of WT and Bgn KO mice by quantitative RT-PCR. n = 4 RT-PCR replicates per mouse, 5 mice per group. d Representative images of α-SMA (green), CD31 immunostaining (red), and DAPI nuclear staining (blue) in E0771 tumors from WT and Bgn KO mice. Scale bar = 100 μm.e Quantification of α-SMA + fibroblasts. The area of α-SMA + fibroblasts was analyzed by excluding α-SMA + pericytes from CD31+ blood vessels. f α-SMA mRNA expression in NIH3T3 cells treated with recombinant biglycan by quantitative real-time RT-PCR. g Cd4, h Klrb1c, i Cd27, j Adgre1, and k Cd8 mRNA expression by quantitative RT-PCR in E0771 tumor tissues from WT and Bgn KO mice. NS, not significant. n = 4 RT-PCR replicates per mouse, 4–5 mice per group. l, m CD8 + CD45+ T cell analysis in E0771 tumors of WT and Bgn KO mice by flow cytometry. l Representative data and m the percentage of CD8 + CD45+ T cells/CD45+ T cells. n = 5 mice per group. n, o Representative images of CD8+ T cells in E0771 tumors from WT and Bgn KO mice. o in tumor center, Scale bar = 100 μm. p Quantification of CD8+ T cells in tumor center. *p < 0.05, **p < 0.01, ***p < 0.001. Significance was determined by two-tailed Student’s t test