Ab272165

Nematodes were synchronized and treated for 1 day with or without 50‐mM metformin starting at L1/L2 or L4 larvae stage. After worms were homogenized in liquid nitrogen, the homogenate was lysed on ice for 60 min in lysis buffer (BioTeKe). Total proteins were loaded (40 μg per well) and separated on a 10% SDS polyacrylamide gel. Proteins were then transferred to Immobilon‐PSQ transfer PVDF membranes (Millipore, Bedford, MA). Primary antibodies against H3K4me3 (1:1,000 dilution; Abcam, ab8580), H3K9me3 (1:1,000 dilution; Abcam, ab8898), H3K27me3 (1:1,000 dilution; Abcam, ab272165), H3K36me3 (1:1,000 dilution; Abcam, ab9050), H3(1:2,000 dilution; Abcam, ab1791), p‐AMPKa (1:1,000 dilution, CST, 2535), p‐S6K (1:1,000 dilution, CST, 9205), S6K(1:1,000 dilution, CST, 2708), p‐mTOR (1:1,000 dilution, CST, 2971), mTOR (1:1,000 dilution, CST, 2972), and beta‐actin (1:1,000 dilution; Abcam, ab227387) used. The secondary antibody was a peroxidase‐coupled anti‐rabbit IgG (1:20,000 dilution; Abmart). The blots were developed using SuperSignal chemiluminescence substrate (Pierce). Band intensities were measured using ImageJ software.

Western blot analysis was implemented using the primary antibodies, rabbit anti‐EZH2 (ab186006, 1:2500, Abcam), rabbit antitrimethylation of H3K27 (H3K27me3) (ab272165, 1:1000, Abcam), rabbit anti‐KLF2 (PA5‐40591, 1:1000, Invitrogen) and rabbit anti‐BIRC5 (10508–1‐AP, 1:1000, Proteintech) as well as the secondary antibody working solution, HRP‐labelled goat antirabbit IgG (ab6721, 1:20 000, Abcam) or goat antimouse IgG (ab6789, 1:20,000, Abcam).¹⁴ Mouse anti‐β‐actin (ab8226, 1:5000, Abcam) served as the internal reference.