Polyclonal rabbit anti trip 1

Total cell lysates were prepared (unless specified in figure legend), and proteins were separated by SDS-PAGE, transferred to PVDF membranes and detected with ECL from GE Healthcare Biosciences (Piscataway, NJ) as previously described [22 (link)], using polyclonal rabbit anti-TRIP-1 from Abcam (1:1000), mouse monoclonal antibodies against smooth muscle actin (1:1000) and tubulin (1:10000) from Sigma-Aldrich and P-Ser473-Akt (1:1000), Smad3 (rabbit monoclonal C67H9, 1:1000), Cleaved Caspase-3 (rabbit monoclonal D175, clone 5A1E, 1:1000 using nitrocellulose 0.2 μm pore size instead of PVDF membranes) from Cell Signaling Technologies, and anti-actin (polyclonal goat I-19, 1:1000) from Santa Cruz Biotechnologies (Santa Cruz, CA). Western-blots were developed using ECL (chemiluminescence), and densitometric scans were quantified using Alpha-Innotech and AlphaEaseFC Imaging Software (Protein Simple, Santa Clara, CA).

Total lung or cell lysates were prepared using previously described methods (Navarro et al. 2009; Perez et al. 2011; Nyp et al. 2014). Western blots were developed using ECL (chemiluminescence) with the following antibodies: polyclonal rabbit anti‐TRIP‐1 (1:1000, Abcam); mouse monoclonal antismooth muscle actin (1:1000) and tubulin (1:10,000) (Sigma‐Aldrich); rabbit monoclonal antiphosphorylated Akt (1:1000) and rabbit polyclonal anti‐Akt (1:1000) (Cell Signaling); mouse monoclonal anti‐E‐cadherin (1:1000, BD Biosciences); and goat polyclonal antiactin (1:2000, Santa Cruz Biotechnologies).