Figure 2.1. Primers for quantification of relative mtDNA copy number per cell.
Primers used for quantification of mtDNA copy number (4kbmtF and 4kbmtR) and nuclear genome copy number (PECAMF and PECAMR). Standard dilution curve, as assayed on whole cell MEF DNA prep, for 4kbmt (A) and PECAM (B) primer sets shows linearity through 1000-fold dilution range.
Figure 2.2. Complete loss of fusion leads to mtDNA depletion. (A) Analysis of
mtDNA:nDNA in WT MEFs and cells lacking Mfn1, Mfn2, both mitofusins, or Opa1 demonstrate that severe loss of mtDNA only occurs with complete ablation of fusion, as in Mfn-null and Opa1-null cells. Infection of Mfn-null cells with either Mfn1 or Mfn2 (C) leads to partial rescue on mitochondrial morphology and to a rapid, sustained increase in mtDNA copy number to that of WT MEFs. In (C), red immunostaining is HSP60 for labeling of mitochondria, and green is for myc, to which Mfn1 and Mfn2 are attached. I.
WT; II. Mfn-null; III. Mfn-null + Mfn1-myc; IV. Mfn-null + Mfn2-myc.
Figure 2.1.
1:10 1:100 1:1000 1:10000 10
15 20 25 30
PECAM Primer Standard Curve
Dilution
Ct Value
1:10 1:100 1:1000 1:10000 10
15 20 25 30
4kbmt Primer Standard Curve
Dilution
Ct Value
A B
Figure 2.2.
I. II.
III. IV.
A
B
C
I II
III IV
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C h a p t e r 3
GENOME-WIDE ANALYSIS REVEALS COATING OF THE MITOCHONDRIAL GENOME BY TFAM
Yun E Wang1, Georgi K Marinov1, Barbara J Wold1, David C Chan1,2
1Division of Biology, California Institute of Technology, Pasadena, CA
2Howard Hughes Medical Institute, California Institute of Technology, Pasadena, CA
Portions of this chapter have been published in PLoS One (Wang, Marinov et al. 2013).