Western lightning plus ecl enhanced chemiluminescence reagent

Stressed P0 parents (at day 2 adulthood) and the unstressed F1 descendants (at day 1 adulthood) were washed by M9 buffer, and frozen in liquid nitrogen. Worm extracts were lysed in sample buffer (2% SDS, 10% glycerol, 5% β-mercaptoethanol, 62.5 mM Tris-HCl pH 6.8, 0.001% bromophenol blue), followed by sonication. After centrifugation, supernatants were collected and boiled for 10 min. Samples were resolved by SDS–polyacrylamide gel electrophoresis (14% polyacrylamide gels) and subjected to immunoblotting according to a standard protocol using primary antibodies (anti-Histone H3 (Abcam ab1791, 1:1,000), anti-Histone H3K4me3 (Abcam ab8580, 1:1,000) and an anti-rabbit secondary antibody (GE Healthcare NA9340, 1:10,000)). Signals were detected with Western Lightning Plus-ECL Enhanced Chemiluminescence reagent (PerkinElmer). Band intensities were measured using ImageJ software. All uncropped western blots can be found in Supplementary Fig. 8.