Work flow chart, linearized map of mtDNA control region, 2-step PCR amplification for target genes and circulating CD34 clones. (A) Flow chart for mtDNA analysis of single circulating CD34 cells and granulocytes (described in “Materials and methods”). Evidence of mtDNA heterogeneity was further confirmed by reamplification of the original lysate. (B) Linearized map and function location of mtDNA control region between nucleotides 16024 to 16569 and 1 to 576 (D-loop) and 2-step PCR amplification of control region (1.12kb) and CO1 (0.91 kb) and Cytb (1.39 kb) genes; HV1 (hypervariable segment 1, nucleotides 16024-16383), HV2 (hypervariable segment 2, nucleotides 57-372), OH (H-stand origin, nucleotides), CSB (conserved sequence block), mt5 (control element), mt3L (L-strand control element), TAS (termination-associated sequence), PL (L-strand promoter), PH1 (major H-strand promoter), TFB (mitochondrial transcription factor binding site), mt4H (H-strand control element), and mt3H (H-strand control element). The homopolymeric C tracts located on HV1 (nucleotides 16184-16193; 5CT4C) and HV2 (nucleotides 303-315; 7CT5C). (C) The number and morphology of circulating CD34 clones after 5-day suspension culture. Each picture shows a different grade: (i) less than 5 cells per well (grade 1), (ii) between 6 and 10 cells per well (grade 2), (iii) 11 to 20 cells per well (grade 3), (iv) more than 21 cells per well (grade 4). Original magnification for panels Ci-Ciii, × 200; for Civ, × 100.