The western blotting analysis was carried out using
Immobilon-P polyvinylidene difluoride membranes (Millipore, 28820 Single Oak Drive, Temecula, California 92590, USA) as described in previous studies 42 (
link),43 (
link). The cell lysates were separated on sodium dodecyl sulfate-polyacrylamide gels and the western blotting analysis was carried out with following antibodies against CSMD1,
Cyclin D1 (Santa Cruz Biotechnology), Snail (Abgent Inc, CA, USA),
E-cadherin,
Vimentin,
Twist, Zeb1 (Cell Signaling Technology, MA, USA), iNOS,
NFκB-p65,
NFκB-p50 (Abcam, Cambridge, CB4 0FL, UK), and
c-MYC (Sigma Chemical Co., MO, USA), respectively. β-actin (TaKaRa Co. Ltd, Kusachi, Japan) was used as an internal control. The density of band was measured using the ChemiDoc™ XRS
+ System (1000 Alfred Nobel Drive, Hercules, California 94547, USA) equipping with Image-Pro Plus software and Epson color image scanner. The data was normalized to the β-actin.
Chen X.L., Hong L.L., Wang K.L., Liu X., Wang J.L., Lei L., Xu Z.Y., Cheng X.D, & Ling Z.Q. (2019). Deregulation of CSMD1 targeted by microRNA-10b drives gastric cancer progression through the NF-κB pathway. International Journal of Biological Sciences, 15(10), 2075-2086.