D4q4z

Whole protein extracts were generated from myoblast cell cultures using an ultrasonic sonicator with a MS73 tip (Bandelin Sonopuls). The proteins were separated by SDS gel electrophoresis using 4–15% TGX gels (BioRad #456–8087). Western blotting was performed using the Trans-Blot^®; TurboTM system (BioRad). Proteins were transferred to low fluorescent PVDF membranes (part of Trans-Blot^®; TurboTM RTA Transfer Kit #170-4274). Membranes were blocked with 5% BSA or 5% skim milk in 1xTBS/0, 1% Tween^®; 20. Following primary antibodies were used: lamin A/C 4A7 (provided by Glenn E. Morris), lamin B1 (D4Q4Z, Cell Signaling) p16INK4A (ab108349, Abcam), p21 (Cell Signaling #2947). For quantification mouse antiGAPDH (Milipore #MAB374) or rabbit antiGAPDH (Cell Signaling GAPDH (D16H11) XP #5174) were used. As secondary antibodies we used donkey anti-mouse IRDye 680RD, donkey anti-mouse IRDye 800CW, donkey anti-rabbit IRDye 680RD and donkey anti-rabbit IRDye 800 CW. All western blot images were obtained using a Licor FC. Quantification was done using the Licor ImageStudio Software. Western blots were repeated at least two times to confirm the results.

Antibodies to CHCHD10 (1:500, 25,671-1-AP, Proteintech, Rosement, IL, USA), M2 (1:1,000, F3165, Sigma-Aldrich), TOM20 (1:1,000, FL-145, Santa Cruz Biotechnology, Dallas, TX, USA), Drebrin (1:500, ab12,350, Abcam, Cambridge, MA, USA), Synaptophysin (1:500, S5768, Sigma-Aldrich), TDP-43 (1:500, MABN45, Merck Millipore, Darmstadt, Germany), LaminB1 (1:1,000, D4Q4Z, Cell Signaling, Danvers, MA, USA), Actin (1:3,000, A5316, Sigma-Aldrich), Actin (1:1,000, sc-1,616, Santa Cruz, Dallas, TX, USA), tRFP (1:1,000, AB233, Evrogen, Moscow, Russia), horseradish peroxidase-linked secondary antibodies (1:5,000, Jackson ImmunoResearch, West Grove, PA, USA), and fluorescently labelled secondary antibodies (1:1,000, Invitrogen) were obtained from the indicated sources.