Figure 3

Expression of mtDNA protein-coding genes is upregulated in BCC and SCC CAFs. (A) qPCR analysis for the expression of mtDNA-encoded genes in BCC CAFs (upper panel) and SCC CAFs (bottom panel), expressed as fold change over isogenic NFs (red dotted line). Color codes indicate the ETC complexes to which the encoded proteins belong to, rRNAs and the Humanin micropeptide. n = 3 biological replicates were used for each graph. Statistical analysis was performed with an unpaired Wilcoxon t test for BCC CAFs and with a paired Mann–Whitney t test for SCC CAFs (the asterisk indicates a comparison surviving Benjamini–Hochberg False Discovery Rate correction). (B) WB analysis for the protein levels of MTCO2 in BCC and SCC CAFs. Upper panel: representative WBs. Arrowhead indicates MTCO2, asterisk designates a cross-reactive unspecific band. Bottom panel: quantitation of MTCO2 expression in n = 3 biological replicates, expressed as fold change over NFs. Statistical analysis was performed with a Wilcoxon t test. (C) Oxygen consumption rates (OCR) at basal and stimulated (maximal) ETC activity in NFs, BCC and SCC CAFs. n = 3 biological replicates were used and analyzed by Kruskal–Wallis test with Dunn’s post hoc test. (D) OCR in basal and maximal conditions in three patient-matched NFs and SCC CAFs pairs analyzed by repeated measures two-way ANOVA with Sidak post-hoc test. (E) Extracellular acidification rates (ECAR) at basal and maximal ETC activity in NFs, BCC and SCC CAFs. Results relative to n = 3 biological replicates and analyzed by repeated measures 2-way ANOVA with Sidak post-hoc test. (F) Quantification of the intensity of Mitotracker Red staining normalized for cell count. Data are expressed as fold change over NFs and derive from n = 3 biological replicates. Analyzed by Kruskal–Wallis test with Dunn’s post hoc test.