Replication Data for: RT-PCR for CDKN1A of "Replicative Senescence in Mesenchymal Stem Cells: An In Vitro Study on Mitochondrial Dynamics and Metabolic Alterations"

Total RNA was isolated from bMSCs (4 × 104 cells/well) using the PureLink RNA Mini Kit (12183018A, Invitrogen, Thermo Fisher Scientific) following the manufacturer’s instructions. cDNA was synthesized in a final reaction volume of 20 µL using the High-Capacity cDNA Reverse Transcription Kit with RNase Inhibitor (4368814, Applied Biosystems, Thermo Fisher Scientific). Quantitative RT-PCR was performed on the CFX96 Real-Time PCR Detection System (Bio-Rad) using TaqMan Gene Expression Assays (Life Technologies, Monza, Italy) for target genes: Cyclin Dependent Kinase Inhibitor 1A (CDKN1A) (Hs003555782_m1) which encodes p21 and mitochondrially encoded NADH dehydrogenase-1 (MT-ND1) (Hs02596873_s1), with GAPDH (Hs99999905_m1) serving as the internal reference gene. Gene expression levels were determined using the 2−ΔΔCt method.