Biotinylated goat anti rabbit secondary antibody

The antibodies used are ERK, ERK pT202/pY204, Akt, Akt pS473, Src, Src pY416, cleaved caspase-3 (Cell Signaling), FAK (BD Transduction Laboratories), FAK pY397 (Millipore), Ki-67 (Novocastra Laboratories), α1(IV) (Abgent), α2(IV) and CD31 (Abcam), α5(IV) (rabbit ployclonal antibody from Proteintech (western blot) and rat monoclonal antibody clone b14 (immunostaining) provided by Dr. Yoshikazu Sado, Shigei Medical Research Institute [36 (link)]), DDR1, phosho-tyrosine (pY99), ubiquitin (Santa Cruz), MEK1 (Abmart), Actin (Sigma-Aldrich), biotinylated goat anti-rabbit secondary antibody (Zymed), Alexa Fluor 555/488 conjugated anti-mouse, rat or rabbit IgG secondary antibodies (Invitrogen). Expression level of integrins on A549 cell surface was determined by immunofluorescence flow cytometry with anti-β1 (Thermo Scientific Pierce), α1, α2 and α11 (Santa Cruz) integrin antibodies as described [37 (link)]. Cycloheximide, MG132 and NH₄Cl were purchased from Sigma-Aldrich.

Histopathological analysis was performed as described before^{25 (link)}. Briefly, mice were sacrificed and lung tissues were inflated and fixed in PFA, embedded in paraffin or frozen in optimal cutting temperature compound (OCT) and sectioned for hematoxylin and eosin (H&E) staining. Immunohistochemical and immunofluorescence analyses were performed as described^{34 (link)}. Antibodies against the following proteins were used: KI-67 (Novocastra Laboratories Ltd), CDC42 (Abcam), ZO1 (33-9100, Zymed); PAR6 (sc-14405, Santa Cruz), Occludin (353197a Zymed), Phalloidin (R415 Invitrogen), CCSP (Santa Cruz), SP-C (AB3786, Chemicon), Beta Galactosidase (ab9631, Abcam), Biotinylated goat anti-rabbit secondary antibody (ZYMED company), Alexa Fluor 555 or 488 conjugated anti-mouse, rat or rabbit IgG secondary antibodies (Invitrogen).