Anti nf1

The western blot analysis involved the use of the following antibodies at specific dilutions: anti-NF1 (1:1000, Proteintech), anti-YAP1 (1:4000, Proteintech), anti-Ubiquitin (1:20000, Proteintech), and anti-β-actin (1:50000, ABclonal). Per the manufacturer's instructions, the NF1 samples were analyzed using an automated capillary-based size sorting system (Wes, ProteinSimple) [30 (link), 31 (link)]. Data analysis was carried out using the built-in Compass software (ProteinSimple).

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).