F1804 1mg

Antibodies against H3 (1: 5000; ab1791), and H3pT11 (1:3000; ab5168) were purchased from Abcam; antibody against FLAG M2 (1:3000; F1804-1MG) was obtained from Sigma-Aldrich; antibodies against Myc (1:5000; 60003-2-1 g), PHGDH (1:3000; 14719-1-AP), GAPDH (1:10000; 10494-1-AP), PSAT1 (1:3000; 10501-1-AP), PSPH (1:3000; 14513-1-AP), PKM1 (1:2000; 15821-1-AP), p21 (1:3000; 10355-1-AP), PCAF (1:3000; 13983-1-AP), SIRT1 (1:2000; 13161-1-AP), 6×His (1:5000; HRP-66005), GST (1:5000; HRP-66001), beta-actin (1:10000; 20536-1-AP), and goat polyclonal anti-mouse IgG (1:5000; SA00001-1) were obtained from proteintech; antibodies against PKM2 (1:3000; 4053 S), p300 (1:5000; 54062), and ATF4 (1:1000; 11815) were purchased from Cell Signaling Technology; antibodies against mouse CDKN1A/p21 (1:3000; A11454), SOD1 (1:2000; A0274), SOD2 (1:1000; A19576), Caspase 3 (1:2000; A11040), PPARγ (1:1000; A11183), PGC1α (1:1000; A220995), and mouse PKM2 (1:5000; A19102) were purchased from Abclonal; antibody against SIRT6 (1:2000; 200499-6C9) was purchased from ZENBO; antibody against PKM2 K433ac (1:500) was custom-made in Abclonal. Antibody against PKM2K305ac was a gift from Dr. Qunying Lei (Fudan University). The specificity of the custom-made antibodies was confirmed by dot blots with peptides or immunoblots with cell extract of corresponding mutants.

Cells were fixed with 4% (wt/vol) paraformaldehyde in PBS (pH 7.4) for 10 min followed by permeabilization with 0.2% (wt/vol) Triton X-100 in PBS before incubation with primary antibodies against Tom20 (rabbit polyclonal, 1:500; Santa Cruz, SC11415) and Flag (mouse monoclonal, 1:100; Sigma-Aldrich, F1804-1MG) for 90 min in 3% BSA, 0.02% Tween-20 in PBS at room temperature. Primary antibodies were labeled with either Alexa Fluor488-conjugated antimouse-IgG or Alexa Fluor568-conjugated antirabbit-IgG secondary antibodies (A-11001 and A-11011; Thermo Fisher Scientific, respectively). Hoechst 33,258 (1 μg·ml⁻¹) was used to stain nuclei. Confocal microscopy was performed using a Leica TCS SP8 confocal microscope (405, 488, 552, and 647 nm; Leica Microsystems) equipped with HyD detectors. Z-sectioning was performed using 300-nm slices and combined. All images were processed using ImageJ (59 (link)).

Antibodies against H3 (1: 5000; ab1791), H3pT11 (1:3000; ab5168) and SETX (1:3000; ab243904) were purchased from Abcam; antibody against FLAG M2 (1:3000; F1804-1MG) was obtained from Sigma-Aldrich; antibodies against Sir2 (1:500; sc-6667) and LC3 (1:1000; sc-398822) were purchased from Santa Cruz Biotechnology; antibodies against Myc (1:5000; 60003-2-1 g), GAPDH (1:10000; 10494-1-AP), beta-actin (1:10000; 20536-1-AP), GFP (1:5000; 66002-1-Ig) and goat polyclonal anti-mouse IgG (1:5000; SA00001-1) were obtained from Proteintech; antibody against Rif1 (1:3000; 95558 S) was purchased from Cell Signaling Technology; antibodies against CDKN1A/p21 (1:3000; A11454), and PPP1CA (1:5000; A12468) were purchased from Abclonal; antibody against CBP (1:3000; abs130593) was purchased from Absin. Antibody against Glc7 (1:5000) was custom-made in Abclonal and its specificity was confirmed by dot blots with Glc7 peptide (Supplementary Fig. S11b). H3pT11 was validated for ChIP assay in H3T11A mutant and PKM2 knockdown HUVECs (Supplementary Fig. S11c, d).

Proteins were separated by SDS–PAGE and transferred to nitrocellulose membranes. In most cases, the protein content was adjusted to 25 μg of total protein per lane using a Nanodrop^® Spectrophotometer ND-1000 to quantify the protein concentration of the samples. Immunoblotting was carried out as previously described^{90 (link)} using a monoclonal antibody against FLAG (1:5000) (Sigma F1804-1MG)) or a polyclonal antibody against 6xHis tag (1:10,000) (Rockland 600-401-382). The secondary antibody Goat anti-Rabbit Ig-HRP (Bio-Rad 172-1019) was added at a 1:20,000 dilution.

The antibody against ASH2L used for Western blots was from Bethyl (A300-489; lot 2) and the one used for immunocytochemistry was from Cell Signaling (CLS D93F6; lot 1). The antibodies against H3K4me3 (Ab8580; lot GR3264593-1), H3K9me3 (Ab176916; Lot GR2318257-2), H3K27me3 (Ab192185; lot GR3264827), and phospho-ATM at serine 1981 (Ab81292; lot GR217573-6) were purchased from Abcam. The antibody against phospho-H2A.X at serine 139 was acquired from Milipore (5636; lot 2554898), and the one against beta actin from Cell signaling (CLS4970; lot 14). Our anti FLAG antibody was purchased from Sigma (F1804-1MG; lot SLGB5673V). The FITC-labeled anti-BrdU antibody was purchased from Thermo Fisher Scientific (11-5071-42, lot 4315462)