Mouse anti pcna

Western blots were performed as described previously^{82 (link),169 (link)}. The following antibodies were used:
Rabbit anti-UHRF1 (polyclonal; 1:250; Cell Signalling, D6G8E), mouse anti-alpha-Tubulin (monoclonal; 1:500; Sigma, T9026), rabbit anti-H3 (polyclonal; 1:1000; Abcam, ab1791), mouse anti-GFP (monoclonal; 1:1000; Roche), mouse anti-FLAG M2 (monoclonal; 1:1000; Sigma, F3165), rabbit anti-xDNMT1 (polyclonal;^{82 (link)}), rabbit anti-xUHRF1 (polyclonal;^{82 (link)}), rabbit anti-USP7 (polyclonal; Bethyl Lab., A300-033A), rabbit anti-H3 (polyclonal; Abcam, ab1791), rat anti-TET1 (monoclonal; 1:10;^{176 (link)}), rat anti-alpha-Tubulin (monoclonal; 1:250; Abcam, ab6160). goat anti-rat HRP (polyclonal; 1:1000; Jackson ImmunoResearch), goat anti-rabbit HRP (polyclonal; 1:1000; BioRad), mouse anti-xCDC45 (monoclonal;¹⁷⁷ ), mouse anti-xRPA2 (monoclonal;^{178 (link)}), and mouse anti-PCNA (monoclonal; Santa Cruz, sc56). Uncropped and unprocessed scans of blots can be found in the Source Data file.

Hairpin Py–Im polyamides 1 and 2 were synthesized on solid phase Kaiser oxime resin using previously published protocols (18 (link)). Gemcitabine, etoposide, hydroxyurea (HU) and doxorubicin were purchased from Sigma-Aldrich, as were all other reagents unless otherwise noted.
Antibodies purchased from Santa Cruz Biotech were: mouse anti-PCNA, anti-Chk1, anti-RPA2, anti-Rad17, anti-FANCD2, goat anti-ATR and rat anti-BrdU (CldU cross-reactivity). Antibodies purchased from Bethyl were: rabbit anti-H2AX, anti-MCM2, anti-MCM2pS108 and anti-RPA2pS4/S8. Antibodies purchased from Abcam were: rabbit anti-FANCD2, anti-MCM2pS108, anti-Rad9, anti-RPA2pS33, anti-Chk2 and anti-H2AXpS139. Antibodies purchased from Cell Signaling Technologies were: rabbit anti-ATMpS1981, anti-Rad17pS645, anti-Chk1pS345, anti-Chk1pS317 and anti-Chk1pS296. Rabbit anti-Chk2pT68 was purchased from Millipore. Rabbit anti-ATM was purchased from Calbiochem. Rabbit anti-ATRpT1989 was a gift of Prof. Lee Zou.

The following antibodies were used: Mouse anti-αSMA-Cy3 (1:1000, Sigma-Aldrich, C6198); Rat anti-CDH1 (1:500, Santa Cruz, sc-59778); Goat anti-KCNJ13 (1:50, Santa Cruz, C19); Rabbit anti-SOX9 (1:400, Millipore, AB5535); Sheep anti-GM130 (1:50, R&D systems, AF8199); Rabbit anti-Ki67 (1:400, Cell Signaling Technologies, #9027); Rabbit anti Cleaved Caspase-3 (1:600, Cell Signaling Technologies, #9661); Rabbit anti KRT5 (1:1000, Abcam, ab53121); Goat anti-CC10 (1:200, Santa Cruz, T-18); Rabbit anti-NKX2.1 (1:400, Santa Cruz, H-190); Rabbit anti-SFTPC (1:400, Millipore, AB3786); Mouse anti-PCNA (1:400, Santa Cruz, sc-56); Rabbit anti-PH3 (1:400, Millipore, 06-570); Rabbit anti-p-AKT^ser473 (1:200, Cell Signaling Technologies, #4060); Rabbit anti-AKT (1:2000 Cell Signaling Technologies, #9272), and Rabbit anti-GAPDH (1:3000, Cell Signaling Technologies, #2118).

Proteins were extracted using RIPA buffer (Solarbio, Beijing, China, R0020) according to the manufacturer’s protocol. The concentration of protein was measured by BCA kit (Takara, Japan, AI90451A). Thirty micrograms of total proteins from each sample were separated by polyacrylamide gel electrophoresis and transferred onto poly-vinylidene fluoride (PVDF) membranes (Millipore, Billerica, MA, USA). The blot-transferred membranes were blocked with 5% fat-free dry milk for 2 h and incubated with primary antibodies: mouse anti-PCNA (1:500; Santa Cruz Biotechnology, PC10), mouse anti-CDK1 (1:500; Santa Cruz Biotechnology, AN21.2), mouse anti-CDK2 (1:500; Santa Cruz Biotechnology, D-12), and mouse anti beta-ACTIN (1:2000; CWBIO, Beijing, China, CW0096) on a shaker at 4 °C overnight, followed by incubation with horseradish peroxidase (HRP)–conjugated anti-mouse IgG (1:2000; Millipore, AP192P). Protein bands were visualized and captured by a Bio-Rad Chemidoc XRS using a Western Bright ECL Kit (Bio-Rad, Berkeley, CA, USA). The protein bands were hand-drawn for 3 times and calculated the average intensity by Image-Pro Plus. The ratio of the target protein to the corresponding intensity value of β-actin was obtained for analysis. Experiments were repeated three times.

The primary and secondary antibodies were purchased from commercial sources as follows: Mouse anti-FTO (Santa Cruz Biotechnology, sc-271713), Rabbit anti-Bax (CST, 2772), Rabbit anti-Bcl-2 (CST, 2876), Mouse anti-PCNA (Santa Cruz Biotechnology, sc-56), Mouse anti-Ki67 (Santa Cruz Biotechnology, sc-23900), Rabbit anti m⁶A (Synaptic Systems, 202003), Rabbit anti-Actin (Sigma Aldrich, A2547), Mouse anti Tubulin (Santa Cruz Biotechnology, sc-365791). HRP conjugated goat anti-rabbit IgG (CWbio, CW0156) and HRP conjugated goat anti-mouse IgG (CWbio, CW0102S).