Rabbit anti p fak

Primary antibodies and titers used in this study for immunostaining are as follows: Rabbit anti-Cleaved Caspase-3 (Asp175) antibody (Cell Signaling, 1:200), mouse anti-Elav antibody (Developmental Studies Hybridoma Bank, DSHB, 1:50), mouse anti-Repo (Developmental Studies Hybridoma Bank, DSHB, 1:50), rabbit anti-GFP (GeneTex, 1:200), mouse anti-NimC1 (a gift from Dr István Andó, 1:30)^{24 (link)}, rabbit anti-Eiger (a gift from Dr Chun-Hong Chen, National Health Research institute, NHRI, 1:200), rabbit anti-FasII (a gift from Dr Vivian Budnik, 1:5000)^{42 (link)} and rabbit anti-pFAK (Cell Signaling, 1:200). For western blotting, mouse anti-Aβ42 (6E10) (Covance, 1:5000), rabbit anti-phospho JNK (pTPpY) (Promega, 1:2000), rabbit anti-JNK (Santa Cruz, 1:5000), and rabbit anti-GFP (GeneTex, 1:5000) were used.

RIPA lysis buffer that contained a protease inhibitor cocktail (Beyotime Biotechnology, Shanghai) was utilized to lyse cells and lung tissues for thirty minutes on ice. The samples were centrifuged at a centrifugation rate of 12,000 g for thirty minutes to sediment the debris. A BCA Assay Kit (Solarbio Life Science, Beijing, China) was used in order to assess the protein content. Proteins (30μg) were isolated using 10% SDS-PAGE and put into PVDF membranes. These plots were then subjected to incubation overnight at a temperature of 4°C with rabbit anti-α-SMA (1 : 500; Abcam), rabbit anti-S100A4 (1 : 1000; Abcam), rabbit anti-p-ERK1/2 (1 : 1000; Cell Signaling Technology), rabbit anti-fibronectin (1 : 1000; Abcam), rabbit anti-ERK1/2 (1 : 1000; Cell Signaling Technology), rabbit anti-FAK (1 : 1000; Cell Signaling Technology), rabbit anti-p-FAK (1 : 1000; Cell Signaling Technology), rabbit anti-p-GSK-3β (1 : 1000; Cell Signaling Technology), and rabbit anti-GSK-3β (1 : 1000; Cell Signaling Technology). As an internal control, GAPDH (1 : 10000; Abcam) was employed. After the membranes were washed, they were exposed to incubation for one hour at room temperature with HRP-conjugated secondary antibodies. The ImageJ program (version: 1.46) from the National Institutes of Health, Bethesda, MD, was utilized for the purpose of conducting the densitometric analysis.