P ulk1ser555

Western blotting was used to examine protein expression following methods detailed previously^{21 (link)}. Briefly, primary human cells (HT01, WRN-Kd, WS01 cells) were collected and prepared using 1x RIPA buffer (Cell Signaling, #9806 S) containing protease inhibitors (Bimake, #B14002) and phosphatase inhibitors (Bimake, #B15002). Proteins were separated on 4–12% Bis-Tris gel (ThermoFisher Scientific, #NP0336BOX) and probed with antibodies. Chemiluminescence detection was performed using a ChemiDoc XRS System. Antibodies used were: β-actin (Santa Cruz, #sc-1616), WRN (Santa Cruz, # sc-5629), CD38 (Santa Cruz, # sc-374650), CD157 (R&D systems, #AF4736), CD73 (R&D systems, #AF5795), PAR (TREVIGEN, #4336-BPC-100), PARP1 (Cell signaling, #9542), AMPK (Cell signaling, #5831), pAMPK (Thr172) (Cell signaling, #2535), pULK1 (Ser555) (Cell signaling, #5869), ULK1 (Cell signaling, #6439), p62 (Cell signaling, #39749), Bcl2L13 (ThermoFisher, # PA5–15043), LC3 (Novus, #NB100–2220), PSD95 (Cell signaling, #3450). All other antibodies were obtained from Cell signaling. Gamma adjustment was used to reduce dark background when necessary. Quantification was performed using ImageJ. Unless elsewhere stated, all the 1st antibodies were with 1000× dilution, while 2nd antibodies were with 10,000× dilution. The original western blots were in the Source Data file with molecular markers labeled.