Anti rbp4

List of anti-mouse protein antibodies used: anti-E-cadherin, anti-N-cadherin (both from Proteintech, Chicago, IL, USA), anti-RBP4 (Abcam, Cambridge, UK), and anti-β-actin-horse radish peroxidase (HRP) (Santa Cruz Biotechnology, Inc., Heidelberg, Germany). Equal amounts of protein (50 μg of cell culture lysates) were mixed with 4× Laemmli Sample Buffer (Bio-Rad Laboratories, Hercules, CA, USA). Then, the samples were separated in a 4–20% sodium dodecyl sulfate (SDS)-polyacrylamide gel and transferred to a polyvinylidene difluoride (PVDF) membrane (0.45 µm; Merck Millipore, city, state abbrev, USA). The membranes were blocked for 1 h at room temperature in 5% non-fat dry milk in 0.1% PBS/Tween-20 (PBST). Next, the membranes were washed (3 × 10 min) with 0.1% PBST and then incubated overnight at 4 °C with a primary antibody. After incubation, the membranes were washed (3 × 10 min) with 0.1% PBST and incubated for 1 h with the secondary mouse anti-rabbit immunoglobulin G (IgG)-HRP antibody (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA). The membranes after washing with 0.1% PBST were detected by the ECL method. Chemiluminescence was visualized using Image Station 4000MM PRO (Carestream Health Inc., Rochester, NY, USA). Densitometry analysis of the blots was performed using Carestream MI Software 5.0.6.20 (Carestream Health Inc., Rochester, NY, USA).