Figure 1.

Effects of induced expression of TFAM-stop and TFAM-MycHis and of RNAi knockdown of TFAM expression. Mitochondrial proteins, DNA and RNA were analysed from Flp-In™ T-Rex™ -293 cells stably transfected with the TFAM-stop (a–c) or TFAM-MycHis construct (d–f), induced over the times indicated or from HEK293T cells (g–i) following transfection with siRNAs Si2 and Si4 over the times indicated. In each case, error bars indicate means ± SEs from at least three independent experiments. a.u., arbitrary units. Measurements of mtDNA levels (a, d and g) are arbitrarily normalized to the mean values for uninduced or untreated cells. For cells under TFAM induction, the measurements were made by two independent methods, Southern blotting and Q-PCR and the values plotted for each time point are the means of measurements by the two methods, shown in Supplementary Figures 1 and 2. (b, e and h) show TFAM protein levels normalized to the mtDNA levels shown in (a, d and g), then normalized against the level in uninduced or untreated cells. (c, f and i) show ND3 mRNA levels normalized first against the 5S rRNA loading control, then against the mtDNA levels shown in (a, d and g), then finally against the level in uninduced or untreated cells. Samples of the raw data and compiled data for TFAM protein, mtDNA and RNA levels on which this figure is based, are shown in Supplementary Figures 1–3.