Fig 1: The VCAN pathway regulates tumor cDC1sFor a Figure360 author presentation of this figure, see https://doi.org/10.1016/j.celrep.2022.111201.(A) Schematic showing versican (VCAN)-V1 functional domains and site-specific proteolysis to generate versikine (scissors represent ADAMTS proteolytic cleavage). CS, chondroitin sulphate.(B) Stromal distribution of anti-DPEAAE IHC staining in human lung cancers. DPEAAE constitutes the C terminus of versikine (chromogen, DAB; counterstain, hematoxylin). 10× objective: scale bars, 240 µm; 40× objective: scale bars, 60 µm.(C) Triple IHC staining of human lung cancers (DPEAAE, teal; XCR1, brown; CD8, purple).(D) Distribution of VCAN expression across TCGA carcinomas (gdc.cancer.gov), ordered on the horizontal axis by median VCAN expression.(E) Distribution of cDC1 (BATF3-DC) score across TCGA carcinomas, ordered on the horizontal axis by median measured cDC1 score.(F) Levels of correlation between cDC1 (BATF3-DC) score and VCAN expression across TCGA carcinomas. The ranked median of VCAN expression and measured cDC1 (BATF3-DC) score is shown across the x axis (1, highest; 20, lowest). Significant (q < 0.1) correlations after multiple hypothesis correction are colored red. Error bars represent the standard error of the correlation coefficient measured using Python statsmodels.(G) Generation of Vcan+/- mice through CRISPR-Cas9-based targeting of Vcan exon 3.(H) Mass cytometry of CD45+ cells from WT (LLC implanted into WT recipients, left) and Vcan-depleted tumors (LLCVcanKD tumor cells implanted into Vcan+/- recipients, right).(I) Quantification of frequency (left) and absolute count ratios (cDC1/cDC1+cDC2 and cDC2/cDC1+cDC2) in WT, Vcan-depleted (LLC-EVVcanKD: Vcan+/-), and versikine (Vkine)-rescued (LLC-VkineVcanKD: Vcan+/-) tumors. Data are presented as mean ± SEM. n = 5 for each group. *p < 0.05, **p < 0.01, ***p < 0.001.(J) Representative flow cytometry plots showing cDC1 and cDC2 frequency in WT, Vcan-depleted (LLC-EVVcanKD: Vcan+/-), and Vkine-rescued (LLC-VkineVcanKD: Vcan+/-) tumors (gating according to Figure S1F).In vitro experiments were performed in technical triplicates. In vivo cohort sizes are shown in individual panels. All experiments were reproduced independently at least twice.