To confirm that human TfR1 is present in the Affi-Gel 15-isolated proteins, including human TfR1 as known from the initial MS analyses, immunoblotting was carried out using anti-TfR1 (Abcam, Cambridge, MA, ab-84036, 1 g/mL) and an HRP-linked second antibody (1:5,000 polyclonal rabbit anti-IgG, 656120, Thermo Fisher Scientific) using whole-cell extracts (1520 g) on 8% or 4%20% SDS-PAGE gels, followed by transfer to PVDF membranes
Cinderellas Carriage is our name for this large, thick fleshed selection that looks exactly like the original in old-fashioned illustrations of the magical fairy tale coach
This restoration of metabolic cofactor availability enables enhanced function of NAD+-dependent enzymes including sirtuins, poly(ADP-ribose) polymerases, and various deacetylases that regulate cellular metabolism, stress responses, and aging processes
Finally, to investigate the molecular basis underlying these phenotypic changes, RT-qPCR and Western blot analyses were performed
