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. 2004 Nov 16;32(20):6015–6027. doi: 10.1093/nar/gkh921

Figure 6.

Figure 6

Transient overexpression of TFAM in HEK cells with lowered mtDNA and TFAM content. (A) Levels of TFAM protein after transfection of cells pre-treated with EBr for 4 days. Control sample of untreated cells shows normal level of TFAM (lane 1); untransfected cells harvested after 18 h (lane 2) and cells which were transfected with pKEX-TFAM for 5 h (lane 3), 12 h (lane 4) and 18 h (lane 5) are shown. p, precursor; m, mature protein. (B) Northern blot showing levels of ND1 mRNA, RNA 19, ATPase 6/8 mRNA, pre-ATPase 6/8 and CO I mRNA as well as the signal of the 28S rRNA used for normalization. Each lane contains RNA from pooled cells of three equally treated culture dishes. Control sample of untreated cells shows normal wild-type level of various RNA species (lane 1), and lanes 2–5 show RNA levels in cells pre-treated with EBr for 4 days. Untransfected cells harvested after 18 h (lane 2) and cells that were transfected with pKEX-TFAM for 5 h (lane 3), 12 h (lane 4) and 18 h (lane 5) are shown. These samples were derived from the same plates as the samples shown on the western blot in Figure 5A.