Figure 2
From: c-Fos induces chondrogenic tumor formation in immortalized human mesenchymal progenitor cells
c-Fos expression induces 3H cells transformation. (a) RT-qPCR of c-Fos expression in transduced cells (n = 3). (b) Western blot showing c-Fos expression (n = 3). (c) Quantification of cell doubling time (n = 3). (d) Cell cycle study showing percentage of cells in each cell cycle phase (n = 3, ANOVA with multiple comparison test). (e) RT-qPCR of Cyclin A1 (CCN1) and B-Myb (MYBL2) expression levels (n = 4). (f) “Reactome_Meiotic recombination” Specific gene set enriched in 3H-Fos cells. Data obtained from GSEA pathway study. (g) Confocal images depicting z-stack reconstruction (Red: Phalloidin staining, Blue: DAPI) (n = 3). (h) Representative images of semisolid cultures at day 7, and quantification of MTT reduction by viable cells recovered from cultures (n = 4). (i) Immunofluorescence images of mitochondrial network (Green: Mitochondria, Blue: DAPI) (n = 3). (j) Quantification of Mitochondrial Membrane Potential (MMP) at basal level (n = 5). (k) Real time measurement of OCR during sequential addition of mitochondrial function modulation compounds (O: Oligomycin, F: FCCP, A/R: antimycin A and Rotenone) (n = 5 per each point, multiple t-test). (l) Lactate accumulation measurement after 48 hours cell culture (n = 3). (m) Cell 6-AN drug dose-response assay (n = 3 per each point). (3H-Ø, empty vector transduced cells; 3H-Fos, c-Fos vector transduced cells). (Unless specified, unpaired t-test. *p ≤ 0.05; **p ≤ 0.01; ***p ≤ 0.001).
