Anti nras

Western blotting (WB) was performed according to previous reports [13 (link)]. The primary antibodies used were anti-β-actin (#4970, Cell Signaling Technology, Danvers, MA, USA), anti-NRAS (#10724-1-AP, Proteintech, Chicago, IL, USA), anti-DOHH (#254866, Abcam, Cambridge, MA, USA), anti-HIST1H2AC (#15953-1-AP, proteintech), anti-TAF6 (#ab76922, Abcam), anti-p44/42 MAPK (Erk1/2) (#4695, Cell Signaling Technology), and anti-Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (#9101, Cell Signaling Technology). Each primary antibody was diluted to 1:1000 and incubated overnight. The secondary antibody, diluted to 1:5000, was incubated at room temperature for 1 h. Protein signals were detected using ChemiDoc XRS Plus Systems (Bio-Rad, Hercules, CA, USA) after a 5 min reaction with the ECL substrate (Bio-Rad). Original images of Western blot analysis please see Supplementary File S1.

Adherent cells were washed, and lysates prepared with pre-chilled RIPA Lysis and Extraction Buffer (Thermo Fisher Scientific, Cat#89900) supplemented with 1–2% Halt Protease and Phosphatase Inhibitor Cocktail (Thermo Fisher Scientific, Cat#78440). Around 50 µg protein lysate per sample was resolved by SDS-PAGE and transferred to a 0.45 µm PVDF membrane. Primary antibodies used were as follows: anti-IQGAP1 (1:200; Santa Cruz Biotechnology (SCBT), cat# sc-376021 or 1:2000; Proteintech, cat# 22167-1-AP), PTPN11 (1:200; SH-PTP2 B-1, SCBT, cat# sc-7384 or 1:1000; Proteintech, cat#.20145-1-AP), anti-p42/44 MAP Kinase (1:1000; Cell Signalling Technology (CST), cat# 9102), anti-pan-Ras (1:200; SCBT, cat# sc-166691), anti-NRAS (1:500; Proteintech, cat# 10724-1-AP), anti-RRAS (1:500; Proteintech, cat# 27457-1-AP), anti-NF1 (1:500; Proteintech, cat# 27249-1-AP), anti-phospho-p42/44 MAP Kinase (Thr202/Tyr204) (1:1000; CST, cat# 9101 S), anti-phospho-AKT (1:1000; CST, cat# 9271), vinculin (1:200; SCBT, Cat# sc-73614), anti-PARP (1:1000; CST, cat# 9542), anti-cleaved PARP (1:1000; CST, cat#9541) and anti-β-Tubulin (1:1000; CST, cat#2146).