Fig. 7: MxIHC and digital cytometry shows fibroblast subpopulations are associated with morphological, molecular and immunological features of LUAD tumours.

a Representative images from whole slide mxIHC analysis of LUAD tumours with varying grades (G1 = well differentiated [n = 1], G2 = moderately differentiated [n = 6] and G3 = poorly differentiated [n = 8]), showing H&E staining and point pattern plots representing the spatial distribution of different fibroblast subpopulations (measured by histo-cytometry analysis of mxIHC data). The scale bar represents 4 mm and the black dotted line demarcates the tumour region in each tissue section. b Boxplots showing the relative abundance of fibroblast subpopulations in well/moderately differentiated LUAD tumours compared to poorly differentiated, measured by CIBERSORTx digital cytometry. Wilcoxon signed-ranks test (n = 375 [G1/G2], 248 [G3]; from two independent datasets;^25,53 data from each individual dataset is shown in Supplementary Fig. 7a). c As per b, measured by scRNA-seq. Wilcoxon signed-ranks test (n = 14 [G1/G2], 7 [G3]). d As per b, measured by mxIHC. Wilcoxon signed-ranks test (n = 7 [G1/G2], 8 [G3]). e Boxplots showing the relative abundance of fibroblast subpopulations in LUAD tumours grouped by TP53 mutation status, measured by CIBERSORTx digital cytometry. Wilcoxon signed-ranks test (n = 578 [wt], 374 [mut]; from two independent datasets^25,51, data from each individual dataset is shown in Supplementary Fig. 7d). f Boxplots showing the relative abundance of fibroblast subpopulations in LUAD tumours grouped by molecular subtype (TRU terminal respiratory unit, PP proximal-proliferative and PI proximal-inflammatory). Each datapoint represents an individual patient, measured by CIBERSORTx digital cytometry (n = 1626, from four independent datasets;^25,51,52,53 data from each individual dataset is shown in Supplementary Fig. 7c). g Scatter plot showing Spearman’s correlation between alveolar or myofibroblast abundance and immune cell subpopulation (LM22) abundance, measured by CIBERSORTx digital cytometry (n = 1626; from four independent datasets;^25,51,52,53 data from each individual dataset is shown in Supplementary Fig. 7e). All statistical tests carried out were two-sided and boxplots are displayed using the Tukey method (centre line, median; box limits, upper and lower quartiles; whiskers, last point within a 1.5x interquartile range). Source data for panels b–f are provided in the Source Data file.