Journal: Reproductive Sciences
Article Title: Conservative Hypomethylation of Mesenchymal Stem Cells and Their Secretome Restored the Follicular Development in Cisplatin-Induced Premature Ovarian Failure Mice
doi: 10.1007/s43032-023-01389-4
Figure Lengend Snippet: Effect of 5-Aza-dC on the methylation pattern of cell’s genomic DNA. Cells were left untreated or grown for 24 h in the presence of a concentration range of 5-Aza-dC (2.5, 5, or 10 μM). Genomic DNA was isolated and left undigested (lanes1–4), digested with the methylation insensitive cutter MspI (lanes 5–8), or with the hypomethylation-sensitive cutter HpaII (lanes 10–13) ( A ). B represents the intensity of DNA fractionation analyzed by Image J. MspI digest generated more DNA fractionation. DNAs derived from 5-Aza-dC treated cells (lanes 10–12) showed a relative increase in HpaII -mediated fractionation compared to normally methylated cells (lane 9). M, molecular weight marker (Lambda DNA-Hind III Digest)
Article Snippet: Cisplatin was purchased from Mylan, Viatris, PA, USA (Cat. no. 198547); 5-Aza-dC was from Sigma-Aldrich (St. Louis, USA, Cat no. 2353-33-5); HpaII and MspI restriction enzymes were from Takara, Bio Inc., Japan, (Cat no.1053A and 1150A, respectively); and alkaline phosphatase reagents were from Spectrum Diagnostics, Egypt (Ref. 216-001).
Techniques: Methylation, Concentration Assay, Isolation, Fractionation, Generated, Derivative Assay, Molecular Weight, Marker, Lambda DNA Preparation