Article Extended Data Fig. 8 | PLA2G2D expression in LN cells and characterization of PLA2G2D-inducing signals. ( a , b ) Analysis of published single-cell transcriptome datasets from lung adenocarcinoma ( a ) and breast cancer ( b ) TDLNs, with UMAP showing cellular diversity within the full datasets. CD68 expression is depicted to highlight the LNM population, in which the bulk of the PLA2G2D-expressing cells reside. ( c , d ) Scaled Pla2g2d ( c ) and Timd4 ( d ) RNA levels projected on the single-cell RNA-seq UMAP. ( e ) qRT–PCR analysis of PLA2G2D mRNA in human monocyte-derived macrophages (MDMs) 2 days after the addition of various dilutions of conditioned medium (CM) from T cell cultures activated with anti-CD3 and anti-CD28 antibodies (n = 3 per group). ( f ) Left: PLA2G2D mRNA expression measured by qRT–PCR in MDMs 2 days after exposure to CM from activated T cells in the presence of anti-IFNγ (15 μ g/ml) or anti-TNF (15 μ g/ml) neutralizing antibodies (n = 3 per group). Right: PLA2G2D mRNA expression measured by qRT–PCR in MDMs 2 days following treatment with recombinant IFNγ (60 ng/ml; n = 4), TNF (60 ng/ml; n = 6), or IFNγ (60 ng/ml) plus TNF (60 ng/ml) (n = 3). ( g ) Log 10 P values and Pearson correlation coefficients (R) between expression levels of genes encoding different cytokines and PLA2G2D in various tumour types (data obtained from TCGA). In case of TGFβ (TGFB), type I and III interferons (IFNs), the isoform with highest R was plotted. IL-6=interleukin 6, IL1B= interleukin 1β, TGFB= transforming growth factor β, TNF=tumour necrosis factor α, BRCA=breast invasive carcinoma, LUSC=lung squamous cell carcinoma, LUAD= lung adenocarcinoma, KIRC: kidney clear cell carcinoma, COAD= colon adenocarcinoma, HNSC= Head and neck squamous cell carcinoma, SKCM= cutaneous melanoma. ( h ) Proportions of lymph-node macrophages (LNMs) of total myeloid cells per treatment group and tissue origin. Group sizes (mice): isotype; LN = 4, TDLN = 8, Tumour=7; anti-PLA2G2D; LN = 4, TDLN = 4, Tumour=4; aIFNγ; TDLN = 4, Tumour=4. ( i ) Violin plot of Pla2g2d RNA levels in untreated and anti-IFNγ treated mice. ( j ) Violin plot of Pla2g2d RNA levels in B16F0 and LN6-987AL inoculated mice. Statistical analysis was performed using a two-sided unpaired t-test ( h ). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. (Exact P values in Supplementary Data).