Fig. 4: mIF analysis of tumors from younger and older patients with TNBC and ER+/HER2− breast cancer.
From: Cell populations in human breast cancers are molecularly and biologically distinct with age
a, Representative images of TNBC tumor tissue sections from younger (<45) and older (>70) patients stained for panCK (red) and vimentin (green). Nuclei are stained with 4′,6-diamidino-2-phenylindole (DAPI) (blue). For each age group, the top rows include individual fluorescent channels and the composite image; the bottom rows include the threshold masks for each channel and the overlap mask obtained for the green and red channels. Enlarged images represent composite images where colocalization of vimentin+/panCK+ cells is indicated in yellow; examples of colocalization are indicated by the white arrows. b, Violin plots representing the quantification of vimentin and panCK overlap staining as a percentage of total panCK staining per image (top) and total panCK staining per image (bottom) in each age cohort. Younger cohort n = 27 independent images representing five tumors; older cohort n = 41 independent images representing seven tumors. c, Representative images of ER+/HER2− tumor tissue sections from younger (<45) and older (>70) patients stained for CD31 (red) and COX4 (green). Nuclei are stained with DAPI (blue). For each age group, the top rows include individual fluorescent channels and the composite image; the bottom rows include the threshold masks for each channel and the overlap mask obtained for the green and red channels. Enlarged images represent composite images where colocalization of CD31+/COX4+ cells is indicated in yellow, and examples of colocalization are indicated by the white arrows. d, Violin plots representing quantification of COX4 staining per image (top), CD31 staining per image (middle) and CD31 and COX4 overlap as a percentage of total CD31 staining (bottom) in each age cohort. Younger cohort n = 29 independent images representing five tumors; older cohort n = 42 independent images representing seven tumors. Data for b and d are provided in Source Data for Fig. 4. Two-sided Wilcoxon rank-sum P values are indicated. Some of the violin plot boundaries exceed 100% or go below 0 because of kernel density smoothing; all values are within the 0–100% range. NS, not significant.
