Fig. 1
SDH-deficiency induces phenotypic changes in imCCs and iMEFs. (A) Maximum velocity of SDH catalysis (SDHmax) calculated according to Eq. 3 (n = 20 cells per line). (B) Quantitation of cell doubling time based on Eq. 2 for imCCs and iMEFs (n = 6 wells per line). (C) Average cell diameter for control and SDH-loss imCCs and iMEFs. Data represent average calculated diameter for 6 fields per line. (D) Percentage of detached cells following trypsinization for 5 min compared to a counting control treated with excess trypsin and mechanical force for total detachment (100%, indicated by red line). P-values are calculated using student’s t-test and the number of asterisks indicates degree of significance. (E) Examples of Flowjo-generated cell cycle analysis histograms showing percentage of the indicated cells in each of the indicated cell cycle phases (G1, S or G2) based on DNA content (measured by fluorescence intensity) using a Watson analysis model (n = 50,000 cells per sample). (F) average percentage of cells in each of the indicated cell cycle phases (G1, S or G2) for each control or SDH-loss line shown as mean ± SEM (n = 3 samples per line processed in a single run). RMSD values for the depicted histograms are Sdhb+/+imCCs: 4.94; Sdhb−/−imCCs: 4.59; Sdhc+/−iMEFs: 4.31 and Sdhc−/−iMEFs: 5.77. These values are representative of the replicate samples. Totals do not all reach 100% because of the presence of cells with DNA content not assigned to any of the indicated phases. P-values are calculated using Bonferroni correction for multiple t-tests and the number of asterisks indicates degree of significance