Anti dock1

All the proteins were obtained from AML tissues and cells using the RIPA lysis buffer and were quantified with the Pierce BCA Protein Assay Kit (Thermo Fisher Scientific, USA). SDS-PAGE (8%) was used to separate the protein, and the concentration of loaded protein samples was 40 μg/lane. After performing electrophoresis at constant pressure, the separated protein bands were electro-transferred onto the PVDF membrane (Millipore, USA) with constant current. The membrane was then washed with PBST (0.1% Tween-20/PBS solution) three times and blocked with 5% BSA for 2 h at room temperature. Subsequently, the rabbit polyclonal anti-DOCK1 (Cat#: ab97325, Abcam, USA) and rabbit monoclonal anti-GAPDH (Cat#: EPR16891, Abcam, USA) were employed for primary antibody incubation overnight at 4 °C. HRP-conjugated Goat anti-rabbit IgG H&L (Cat#: ab181602, Abcam, USA) was used for secondary antibody incubation for 2 h at room temperature. After repeated washing with PBST, the blots were visualized using the enhanced chemiluminescence kit (Thermo Fisher Scientific, USA).

Western blot analyses were conducted as reported 17 (link). The antibodies used were: anti-Dock1 (abcam, 1:1000), Vimentin (abcam, 1:1000), E-cadherin (abcam, 1:1000), Snail (abcam, 1:500), p-IκB (CST, 1:1000), IκB (CST, 1:1000) antibody. All experiments were repeated triply. The gray value of western blot were evaluated and quantified using ImageJ software.