Anti dab2

PE/Cy7-tagged CD45 (Cat# 103114), APC-tagged Ly6C (Cat# 128016), FITC-tagged CD66b (Cat# 555724), V450-tagged Ly6G (Cat# 560603), Alexa fluor 700-tagged CD11b (Cat# 557960), anti-human PE/Cy7-tagged CD45 (Cat# 560178), and Anti human CD34 antibody (Cat# 343602) were purchased from BD Biosciences, USA and anti-human PE-tagged IL-28AR antibody (Cat# 337804) was purchased from Biolegend, USA. Anti-Neutrophil Elastase (Cat# ab68672), anti-GRO alpha (CXCL1) (Cat# ab86436), Anti-STAT1 (phosphor S727) (Cat# ab109461), anti-IL28 receptor alpha or IL28R1 (Cat# ab224395), anti-Histone H3 citrunillated (Cat# ab219407), VCAM1 (Cat# ab134047), CD34 (Cat# 8158) and IL28 + IL29 (Cat# ab191426) antibodies were purchased from Abcam, USA. Anti-ICAM-1 (Cat# SC-107), Anti-STAT1 (Cat# 9172T), anti-AKT (Cat# 4685S), anti-AKT2 (Cat# 2964S) and anti-DAB2 (Cat# 12906S) were purchased from Cell Signaling Technologies, USA. Other antibodies used include: Alexa fluor 488-tagged β1 Integrin (Santa Cruz Biotechnology, USA; Cat# sc-374429 AF488), Anti-IL-28A/IFNλ2 (Antibodies online, USA; Cat# ABIN357173), Anti-Ly6G (Antibodies online, USA; Cat# ABIN1854937), IL-29 antibody (Biorbyt, USA; Cat# orb6201), anti-IFNα (Thermo Fisher, USA; Cat# 221001), anti-Myeloperoxidase/MPO (R&D Systems, USA; Cat# AF3667-SP), anti-LCN-2 (EMD Milipore; Cat# AB2267) and anti-Actin (Sigma Aldrich, USA; Cat# A2066).

Slides of proliferating or differentiating K562 cells were prepared using TXT3 Cytospin (Nasco Korea Corp., Seoul, Korea) at 900 × g for 5 min. Slides were rinsed in phosphate-buffered saline (PBS; pH 7.4), fixed with 4.0% paraformaldehyde (PFA) for 10 min, and permeabilized using PBS-T buffer [0.1% Triton X-100 in 1 × PBS, phenylmethylsulfonyl fluoride (PMSF), phosphatase inhibitor cocktail (Roche, P4906845001), and protease inhibitor cocktail (Sigma, P8340)] for 10 min on ice. Primary antibody staining was performed using anti-DAB2 (Cell Signaling, 12906, 1:1000). Secondary antibody staining was performed using Alexa Fluor™ Plus 488 conjugated anti-rabbit IgG (Invitrogen, A32731, 1:500). DNA was counterstained with DAPI (Thermofisher Scientific, 62248). A Zeiss LSM880 confocal microscope was used for imaging.

Slides of proliferating or differentiating K562 cells were prepared using TXT3 Cytospin (Nasco Korea Corp., Seoul, Korea) at 900×g for 5 min. Slides were rinsed in phosphate-buffered saline (PBS; pH 7.4), fixed with 4.0% paraformaldehyde (PFA) for 10 min, and permeabilized using PBS-T buffer [0.1 % Triton X-100 in 1×PBS, phenylmethylsulfonyl fluoride (PMSF), phosphatase inhibitor cocktail (Roche, P4906845001), and protease inhibitor cocktail (Sigma, P8340)] for 10 min on ice. Primary antibody staining was performed using anti-DAB2 (Cell Signaling, 12906, 1:1000). Secondary antibody staining was performed using Alexa Fluor^™ Plus 488 conjugated anti-rabbit IgG (Invitrogen, A32731, 1:500). DNA was counterstained with DAPI (Thermofisher Scientific, 62248). A Zeiss LSM880 confocal microscope was used for imaging.