1. Volume 26, Issue 2, Pages 429-436.e4 (August 2017)
Increased Total mtDNA Copy Number Cures Male Infertility Despite Unaltered mtDNA Mutation Load 
Min Jiang, Timo Eino Sakari Kauppila, Elisa Motori, Xinping Li, Ilian Atanassov, Kat Folz-Donahue, Nina Anna Bonekamp, Sara Albarran-Gutierrez, James Bruce Stewart, Nils-Göran Larsson 
Cell Metabolism 
Volume 26, Issue 2, Pages e4 (August 2017)
DOI: /j.cmet
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2. Cell Metabolism 2017 26, 429-436.e4DOI: (10.1016/j.cmet.2017.07.003)
Copyright © 2017 Elsevier Inc. Terms and Conditions

3. Figure 1 TFAM Overexpression Rescues Infertility Phenotypes
(A) Mating strategy.
(B) Pups per litter.
(C) Relative testis weight.
(D) Relative sperm count.
(E) Sperm motility.
(F) Sperm morphology with representative examples of (a) normal sperm, (b and c) sperm head defects, and (c–e) sperm tail defects (scale bar, 20 μm).
(G) Quantitative assessment of sperm morphology. At least three mice were included for each genotype and more than 300 sperm were assessed from each mouse.
Data are represented as mean ± SEM; ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p <
Cell Metabolism  , e4DOI: ( /j.cmet )
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4. Figure 2
TFAM Overexpression Partially Rescues Structural and OXPHOS Defects of the Seminiferous Epithelium
(A) Hematoxylin and eosin (HE) staining of testis sections of 25-day- (a–d), 6-week- (e–l), and 4-month-old (m–p) mice of the shown genotypes (i–l are the higher magnifications of the red frames in e–h).
(B) COX/SDH staining of testis sections. (e)–(h) are higher magnifications of the red frames of (a)–(d), respectively. Arrowheads indicate the sperm tails; arrows indicate the round spermatids.
(C) Immunohistochemistry of COXI in the testis sections; brown color is the signal of COXI protein. (e)–(h) are higher magnifications of the red frames shown in(a)–(d), respectively. Red circles indicate the round spermatids in the testis sections; arrows indicate the pachytene spermatocytes (scale bar, 50 μm).
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5. Figure 3
TFAM Overexpression Partially Rescues Mitochondrial Cristae Structures in Pachytene Spermatocytes, Spermatids, and Sperm
(A) Mitochondria in later pachytene spermatocytes and round spermatids. The area indicated by a black frame is further enlarged to better visualize mitochondrial morphology (scale bar, 2.5 μm).
(B) Quantification of mitochondrial morphology (normal type, condensed; abnormal types, intermediate and long form) in round spermatids (each genotype included more than three mice and more than five round spermatids were assessed from each mouse).
(C) Representative images of the mitochondrial sheath of sperm from cauda epididymis of the shown genotypes (scale bar, 500 nm).
Data are represented as mean ± SEM; ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p <
Cell Metabolism  , e4DOI: ( /j.cmet )
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6. Figure 4
TFAM Overexpression Reverses Defects in OXPHOS Subunits Caused by mtDNA Mutations
(A) mtDNA copy number in spermatocytes.
(B) Total mutation load in spermatocytes.
(C) Downregulated mitochondrial proteins in PolgAmutTfamWT spermatocytes.
(D) TFAM protein expression level and upregulated mitochondrial proteins in PolgAmutTfamWT spermatocytes. For each genotype, spermatocytes from at least four mice were included.
Data are represented as mean ± SEM; ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p <
Cell Metabolism  , e4DOI: ( /j.cmet )
Copyright © 2017 Elsevier Inc. Terms and Conditions