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. 2017 Feb 2;7:41490. doi: 10.1038/srep41490

Figure 5. Mule is indispensable for the maintenance of mitochondrial biogenesis and bioenergetics.

Figure 5

(A) Immunoblot analysis of master transcription factors regulating FAS and FAO in left ventricular extracts of Mulefl/fl(y);mcm and DKO mice at 7 days post-Tam. Numbers below the individual blots indicate fold-changes in protein expression. (B) Endogenous Myc interacts with gene promoter sequences of Esrrβ, but is not associated with the promoters of Pparα and Pparγ as analyzed by ChIP. Esrrβ, estrogen related receptor, beta. (C) Transcript expression of key factors regulating FAS/FAO, redox homeostasis and mt biogenesis in left ventricular samples of Mulefl/fl(y);mcm mice was determined by RT-qPCR at 4 weeks post-Tam. n = 4. *P < 0.05 versus vehicle. (D) ATP levels in left ventricular tissue extracts from Mulefl/fl(y);mcm mice at 7 d post-Tam. n = 4. (E) Immunoblot analysis of important constituents of the mt electron transport chain participating in oxidative phosphorylation in LV extracts of Mulefl/fl(y);mcm versus DKO mice at 7 days post-Tam. Equal loading of bands was confirmed by reprobing membranes with mt- specific anti-Opa1 antibodies. (F) Myc interacts with gene promoter sequences of OxPhos components of Ndufa4 (CI, complex 1) and Cox10 (CIV, complex 4) as analyzed by ChIP from isolated LV genomic DNA from Mulefl/fl(y);mcm mice at 7 d post-Tam. (G) Mt biogenesis, defined as relative DNA copy number of mt encoded Cytb gene normalized to the copy number of the nuclear gene Npm1, was determined by qPCR of isolated LV mt from Mulefl/fl(y);mcm and DKO mice at 7 d post-Tam. n = 4. (H) Mt capacity, defined as relative mRNA levels of the nuclear gene Ndufa8, a subunit of OxPhos complex I, normalized to Npm1 transcript expression, was determined by RT-qPCR of total RNA isolated from left ventricular tissue of the indicated wild type (−Tam) and mutant strains (+Tam). n = 4. (I) Activity of OxPhos complex I in Mulefl/fl(y);mcm and DKO mice at 7 d post-Tam. n = 4. *P < 0.01 versus vehicle. (J) Activity of OxPhos complex IV in Mulefl/fl(y);mcm mice or DKO animals at 7 d post-Tam. n = 4. *P < 0.05 versus vehicle. Data are means ± s.e.m.