U2OS 3XMyc-EGFP-OMP25 (Control-mitochondria) and 3XHA-EGFP-OMP25 (HA-mitochondria) stable cell lines were generated according to the published protocol.74 Retrovirus-transduced cells were selected with 10 μg/mL blasticidin for 2 weeks, before sorting low GFP expressing populations (bottom 25% ...
but given the high mutation rate of mtDNA and the absence of sexual reproduction, mitochondria still have to cope with Müller’s ratchet, so there must be a mechanism to avoid the transmission of damaged organelles through generations. Despite the high mtDNA copy number in mature oocytes (about...
Cells were transfected with GenJet DNA In Vitro Transfection Reagent version II (SL100489; SignaGen Laboratories) following the protocol suggested by the manufacturers. After 48 hours, cells were sorted using a FACSAria IIU by gating on single-cell fluorescence using a 561-nm laser and 600LP,...
mtDNA copy number in the Mes, as well as between mtDNA copy number and chemerin expression in the Mes, in the group with BMI > 40 kg/m2 without T2DM demonstrates the protective effect of chemerin against the development of IR via the regulation of mtDNA copy number in adipose ...
Due to the adverse influence of aging, an “add-on” or adjuvant medical intervention is commonly given, either alone or in combination, to the standard protocol of IVF to improve outcomes [31, 32]. LH and GH are two substances ubiquitously utilized for this purpose. Involved in the complet...
It has been found that tumor cells undergoing oxidative stress can lead to mitochondrial dysfunction, which is closely associated with reduce of mtDNA copy number [8, 9]. Since Spees et al. has proved that mitochondria or mtDNA can move between cells [30], studies further point out that mito...
(Invitrogen #L3000001) was used for transfection according to the manufacturer’s protocol. Specifically, we used 1 µg DNA and the ratio of p3000 to the 3000 reagent was 2:1. The cells were imaged 2 days after transfection in recording media with the LSM 880 microscope using a 20x/...
Since mtDNA is maternally inherited, exists in a high copy number (1,000–10,000 copies) in each cell, and is rapidly evolving (5–10 times faster than nuclear DNA), sequence poly- morphism of the mtDNA has proved useful in the fields of population and evolution study (Gresham et al ...
DNA was isolated with QIAamp DNA Micro kit (QIAGEN) following the manufacturer’s protocol. For the quantification of mitochondrial DNA copy number a multiplex RT- PCR assay was used according to Phillips et al. (2014). DNA samples (50ng/reaction) were used together with appropriate volumes ...
Mitochondrial function is critical to continued cellular vitality and is an important contributor to a growing number of human diseases. Mitochondrial dysfunction is typically heterogeneous, mediated through the clonal expansion of mitochondrial DNA (mtD