Extended Data Fig. 3: Proteomics analysis of HPFs in co-cultures and CAF features of recipient fibroblasts.
a. PCA of proteomics data from MitoTracker-high and -low and control HPFs. n = 4 co-cultures and sorting experiments. b. Volcano plot showing differentially abundant proteins in sorted MitoTracker-low vs. control HPFs. n = 4 co-cultures per group. Selected CAF marker proteins are in red. c. Volcano plot showing differentially abundant proteins in sorted MitoTracker-high vs. HPFs. n = 4 co-cultures per group. Selected CAF marker proteins are in red. d. Unbiased pathway analysis of MitoTracker-high vs. - low HPFs referenced to Hallmarks of Cancer (left) and WikiPathways (right). e. Gating strategy (top panel) and representative histogram of flow cytometry for Ki67+ cells among RFP-high and -low and control HPFs (bottom panel). Gating was set based on the negative control. f. Percentage of Ki67+ cells among RFP-high- and -low HPFs after co-culture with A431 cells expressing Su9-RFP. n = 3 co-cultures per group. g. qRT-PCR for INHBA, IL6, ACTA2 and COL1A1 relative to RPL27 using RNA from RFP–high and -low HPFs after co-culture with A431 cells expressing Su9-RFP. n = 3 co-cultures per group. h, j. Percentage of Ki67+ cells and transwell migration of primary cutaneous SCC cells (h), SCC13 cells (i) and HaCaT-Ras cells (j) cultured with CM from control or MitoTracker-low or -high HPFs sorted after co-culture with the respective cancer cell lines. n = 3 co-cultures per group. Graphs show mean ± SEM. Unpaired two-sided Student’s t-test (f, g) or two-sided one-way ANOVA with Bonferroni post-host multiple comparison test (h–j) were used to determine statistical significance.