Anti lats2

Anti-LATS2, YAP1, lamin-B, PPD-L1, GAPDH rabbit polyclonal (Abcam, Cambridge, United Kingdom), and anti-actin polyclonal (Santa Cruz Biotechnology) antibodies were used at 1:1,000 dilution for Western blotting. Anti-rabbit polyclonal secondary horseradish peroxidase-conjugated antibodies were used for detection (diluted; 1:2,000). Paclitaxel (TX) was purchased from Sigma (St. Louis, MO). The working stock was diluted in the media at a final concentration of 4μM and further diluted when needed. The human ovarian serous cancer cell line, MCF7 cells were obtained from ATCC (Manassas, VA, United States). These cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) plus 10% fetal bovine serum (FBS) at 5% CO₂ and 37°C.

The protein concentration was measured by the BCA method and quantified. The experimental procedure was carried out according to a conventional Western blotting protocol. The proteins were separated using 10% SDS-PAGE and transferred to a PVDF membrane under constant current of 320 mA. The membrane was blocked with 5% skim milk at room temperature for 1.5 h and with 1:1000 dilutions of anti-LATS2 (Abcam), anti-E-cadherin (Abcam), anti-N-cadherin (Abcam), anti-α-SMA (Alpha-smooth muscle actin) (Abcam), anti-CD9 (Abcam), anti-CD63 (Abcam), and anti-CD81 (Abcam) overnight at 4 °C. The next day, the membrane was washed three times with TBST and incubated with a 1:1000 diluted HRP-conjugated secondary antibody (Abcam) for 1 h at 37 °C. After washing three times with TBST, chemiluminescence was performed using an ECL reagent (Bio-Rad). The band intensity for each protein on the membrane was scanned by a scanner and analyzed by image processing software.