The protocol was modified from Ross (2011). Briefly, fresh testes tissue were embedded in optimal cutting temperature (OCT) compound, cryosectioned into 10-μm-thick sections, mounted on slides, and left to air dry briefly. Freshly prepared buffer A (0.8 mL of 5 mM 3,3′-diaminobenzidine...
A real-time PCR assay was developed to measure total mtDNA copy number (mtDNACN) and mtDNA harboring the 5 Kb “common deletion” to enable calculation of the mtDNA deletion ratio (mtDNADR) in 143 cultured oocytes. Kruskal-Wallis test was carried out to compare the total mtDNACN and the ...
Cells were transfected using the SF cell line 4D-nucleofector X kit (Lonza) with the CN-114 program according to the manufacturer’s protocol. One million cells were transfected with a total of 2,000 ng of plasmid DNA (1,000 ng of each DdCBE monomer). Cells were split 24 h aft...
Cells were transfected using the SF cell line 4D-nucleofector X kit (Lonza) with the CN-114 program according to the manufacturer’s protocol. One million cells were transfected with a total of 2,000 ng of plasmid DNA (1,000 ng of each DdCBE monomer). Cells were split 24 h aft...
A better understanding of molecular pathways involved shall precede the identification of newer diagnostic tools and therapeutic targets. We explored EPOR signaling pathway in Circulating tumor cells (CTCs) and HNSCC cell lines.Methods and Results: By using a validated in-house CTC isolation protocol,...
that followed the manufacturer protocol. That means that we used the dyes within the indicated concentration/timing range. Obviously it is impossible to use exactly the same protocol in every species, because of intrinsic differences (e.g.: different osmolarity, yolk storage, etc.). Protocol optim...
protocol. Full size image To illustrate potential structural consequences of mutations, two examples exhibiting a very high (>80%) variant level and severe alteration of the HRR profile were examined in detail: the variantT11991C, recently also detected in a PCa tissue sample (COSM1132242, [...
The study protocol was approved by the Ethics Committee of Wenzhou Medical University. Genomic DNA from the volunteers was extracted using sodium dodecyl sulfate (SDS) lysis buffer as described previously [8]. Sanger sequencing was performed for the entire mtDNA of each individual on an ABI 3730...
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...
000 × gfor 30 min at 4 °C for production of a supernatant corresponding to the cytosolic fraction. DNA of cytosolic fractions were isolated using QIAQuick nucleotide removal kit (28306, QIAGEN, Valencia, CA) following the manufacturer’s protocol. The copy number of mtDNA was ...