Fig. 3: Transferred cancer cell mitochondria induce a CAF phenotype.
a–c, Volcano plots displaying differentially expressed genes in MitoTracker-low versus control (a), MitoTracker-high versus control (b) and MitoTracker-high versus MitoTracker-low HPFs (c) sorted from n = 3 cocultures with A431 cells. d,e. RNA-seq data from sorted HPFs depicting expression of INHBA, IL6, ACTA2 and COL1A1 (d) or PDGFRA, PDGFRB, S100A4, FAP and CD74 (e) in MitoTracker-high, MitoTracker-low and control groups sorted from n = 3 cocultures. f–h, Comparative analysis of gene signatures in MitoTracker-high versus MitoTracker-low HPFs with published CAF datasets23,28,30, showing similarities of MitoTracker-high HPFs with myCAFs and iCAFs. i, Volcano plot displaying differentially abundant proteins in MitoTracker-high versus MitoTracker-low HPFs sorted from n = 4 cocultures with A431 cells. j, Correlation analysis of gene and protein expression in MitoTracker-high versus MitoTracker-low HPFs. Significantly regulated pathways (q < 0.1) are highlighted (blue, Hallmarks of Cancer pathways; purple, Wikipathways). k, Percentage of Ki67-positive HPFs and relative levels of intracellular ATP and MitoSOX in sorted HPFs (n = 3 cocultures for Ki67 and 6 cocultures for ATP and MitoSOX). l, Percentage of Ki67-positive A431 cells in spheroids cultured with CM from control HPFs and sorted MitoTracker-high and MitoTracker-low HPFs (n = 3 spheroids per treatment group). m, Transwell migration of A431 cells in CM from sorted MitoTracker-low and MitoTracker-high and control HPFs (n = 3 transwell cultures per treatment group). n. Relative colony size of A431 cells plated on dECM from sorted HPFs (n = 3 cultures). Graphs show the mean ± s.e.m. An unpaired two-sided Student’s t-test (k (right and middle),n) or two-sided one-way ANOVA with Bonferroni post hoc multiple comparison test (d,e,k (left),l,m) was used to determine statistical significance. One control value was set to 1.
