Anti gapdh

Western blotting was conducted as described previously (Chen et al., 2017 (link); He et al., 2020 (link)).The following antibodies were used: anti-SVEP1 (1:500) from R&D (MAB97741), anti-GAPDH (1:1000) from Santa Cruz Biotechnology.

Genomic DNA was isolated using the DNeasy Blood and Tissue kit (69506, Qiagen). Unmethylated cytosines were converted to uracil by sodium bisulfite using the EZ DNA Methylation-Gold kit (D5005, Zymo Research, CA). Modified DNA was amplified with primers targeting the CpG sites (cg04087271, cg20619374, and cg06984156) of CDA (Supplementary Table 3). Gel-purified bands were extracted using a Gel Extraction kit (28706, Qiagen) and cloned into the pGEM-T Easy vector (A1360, Promega, WI). Multiple plasmid DNA was isolated using the HTS Plasmid kit (PHTS-30, Core Bio System, Korea), and Sanger DNA sequencing was performed by GenoTech (Daejeon, Korea).
Western blot analysis Western blotting was performed as described^{14 (link)}. Antibodies were diluted in 5% skim milk or 5% bovine serum albumin in Tris-buffered saline containing 0.1% Tween-20. The antibodies and dilutions used were as follows: anti-CDA (SAB1300717, Sigma–Aldrich, 1:500), anti-E-cadherin (1:1000, 3195, CST, MA), anti-N-cadherin (1:1000, 4061, CST), anti-vimentin (1:1000, 5741, CST), anti-GAPDH (1:2000, sc-47724, Santa Cruz Biotechnology, CA; 1:2000, 5174, CST), goat anti-mouse IgG (1:5000, 31430, Invitrogen, CA), and goat anti-rabbit IgG (1:5000, 31460, Invitrogen). Immunopositive bands were detected using an ECL kit (K-12045-D50, K-12042-D10, Advansta, CA) and visualized using a Fujifilm LAS-4000 (Tokyo, Japan).

The mice sacrificed by anesthesia using CO2. Total protein was extracted from mouse liver using protein lysate (RIPA: PMSF = 100:1). The total protein was determined by the BCA protein assay kit (Thermo). Proteins in samples were fully denatured by heating in 4× loading buffer at 100 °C for 10 min, separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and transferred to polyvinylidene difluoride membranes (Millipore, Billerica, USA). Membranes were blocked with non-fat milk and probed with primary antibodies (anti-TfR1, anti-FPN1, and anti-GAPDH) (Santa Cruz). Membranes were then incubated with horseradish peroxidase-conjugated anti-rabbit IgG (Santa Cruz). After washes three times the blots were visualized by enhanced chemiluminescence with chemiluminescence system machine (Clinx Company, ChemiScope 6300). Gray value of Western blot strips was quantified with Image J software (National Institutes of Health).

Viable cells in PBS buffer or cell lysates in RIPA buffer were reduced with DTT in 2x Laemmli buffer and denatured at 100°C. Western blotting was carried out as previously described [41 (link)]. Target proteins were detected using anti-α-tubulin (1:10000, T5168, Sigma‒Aldrich, St. Louis, MO, USA), anti-TRMT112 (1:500, sc-398481, Santa Cruz Biotechnology, Dallas, TX, USA), anti-Cyclin B1 (1:500, sc-245, Santa Cruz Biotechnology, Dallas, TX, USA), anti-Cyclin E (1:500, sc-247, Santa Cruz Biotechnology, Dallas, TX, USA), anti-Cyclin A (1:500, sc-271682, Santa Cruz Biotechnology, Dallas, TX, USA), anti-N6AMT1 (1:1000, CQA1550, Cohesion Biosciences, London, UK), and anti-GAPDH (1:2000, sc-32233, Santa Cruz Biotechnology, Dallas, TX, USA) antibodies. Goat anti-rabbit-HRP (1:10000, 31460, Invitrogen, Carlsbad, CA, USA) and goat anti-mouse-HRP (1:10000, 31430, Invitrogen, Carlsbad, CA, USA) were used as secondary antibodies.