Nad nadh kit

Assays were performed with NAD⁺/NADH kit (Sigma, MO) according to the manufacturer’s instructions. Cell pellets were lysed using a Qiagen Tissue Lyser LT (Qiagen, Germany) at 50 oscillations/s for 3 min in the NAD⁺/NADH extraction buffers (Sigma, MO), the resulting lysate was then used to assay for NAD⁺/NADH quantification with the aid of colorimetric indicators at 450 nm. Spectrophotometric measurements were conducted with an iMark^TM microplate reader (Bio-Rad, CA).

For metabolic assays, fibroblasts were converted for three weeks and 50.000- 100.000 FACS-sorted iNs were plated on Geltrex-coated, white- or black-walled 96-well plates. Four to seven days after re-plating, supernatant was analyzed using colorimetric lactate assays (Biovision, #K607 or Merck MAK064). Attached cells were processed using the Pyruvate Kinase activity assay Kit (Sigma Aldrich, MAK072), the ATP/ADP ratio kit (Sigma Aldrich, MAK135), or the NAD/NADH kit (Sigma Aldrich, MAK037 or Promega, G9071) according to the manufacturers’ instructions. Recording of signals was performed using the Enspire Multimode Plate Reader platform (PerkinElmer).