Extended Data Fig. 2: DC frequency in TME and in vivo CRISPR screening QC.

a, Flow cytometric gating strategy for DC populations in TME. b,c, Quantifications of total cDCs, CD103⁺ cDCs, and CD11b⁺ cDCs in TME from early or late stage (b, B16F10, n = 5 mice; c, MC38, n = 6 mice). d, LAMP3⁺ DC signatures in early- or late-stage cancer patients based on TCGA database. e,i, All the mapped sgRNA hits for the pre-round (e) or major-round (i). Data are merged from 5 (e) or 3 (i) independent experiments. The x-axis indicates sgRNA numbers and y-axis indicates the calculated log2 fold change. f,j, Coverage calculation in each step from pre-round (f) or major-round (j) screening. g,k, Gini-Index analysis for pre-round (g) or major-round (k) screening. h, Minipool gRNAs selection strategy after the pre-round screening. l, Distribution of the enriched sgRNAs for top-hit genes from the major-round screening. m, Western blot analysis of Pde5 expression in BMDCs derived from Pde5^+/+ or Pde5^−/− mice. β-actin was used as control. n, Frequencies of CTV-labelled Pde5^+/+ and CFSE-labelled Pde5^−/− DCs in the TME two days post peritumour transfer (n = 6 mice). Quantifications on right panel were normalized to the mixture ratio before cell transfer. o, Whole-mount tissue 3D reconstitution (left) and cross-section image (middle) of tdLNs from mice co-transferred with Pde5^+/+ and Pde5^−/− DCs. Cells were surface rendered and DC signals were processed as dots. Scale bars, 500 μm. Data quantified from full tissue sections from each mouse (n = 2 mice). p, Representative staining and quantification of CTV-labelled Pde5^+/+ and CFSE-labelled Pde5^−/− mature BMDCs in mouse ear explants (n = 7 lymphatic vessels). Scale bar, 100 μm. Data are representative (b, c, m, n) or merged (e, g, i, k, l, o, p) of at least two to three independent experiments and present as mean ± s.e.m. Statistical analysis was performed using two-tailed Student’s unpaired (b, c) or paired (n, p) t-test or two tailed Mann–Whitney U-tests (d).

Source data