Blocking buffer: 5% NFDM/TBST Primary ab dilution: 1:2000 Primary ab incubation condition: lane 1: bs-60081R+H3K4me3/Q5ser peptides, lane 2: bs-60081R+unmodified peptides, lane 3: bs-60081R, 2 hours at room temperature Secondary ab: Goat Anti-Rabbit IgG H&L (HRP) Lysate: N2a Protein loading quantity: 20 μg Exposure time: 60 s Predicted MW: 15 kDa Observed MW: 15 kDa

Blocking buffer: 5% NFDM/TBST Primary ab dilution: 1:2000 Primary ab incubation condition: 2 hours at room temperature Secondary ab: Goat Anti-Rabbit IgG H&L (HRP) Lysate: HeLa, N2a, BRL, Mouse kidney Protein loading quantity: 20 μg Exposure time: 60 s Predicted MW: 15 kDa Observed MW: 15 kDa

Chromatin immunoprecipitation was performed using non-cross-linked chromatin from 5×10⁶ HeLa cells per IP, with 6 μg of Histone H3 (tri methyl K4, serotonyl Q5) Rabbit pAb（bs-60081R） or 6 μg of normal rabbit IgG (negative control) , and 50 μL of Protein A MagBeads per IP. The immunoprecipitated DNA was quantified by real-time PCR using primers specific for the human GAPDH-CDS, RPL30 exon 3, LDHA, FOXO3a-promoter, FOXO3a-downstream, RAB20, and LDHA-promoter regions. The data are presented as enrichment of each sample relative to the total amount of input at each amplicon.