Studies on Multifunctional Effect of All-Trans Retinoic Acid (ATRA) on Matrix Metalloproteinase-2 (MMP-2) and Its Regulatory Molecules in Human Breast Cancer Cells (MCF-7)
Figure 3
Western blot of MT1-MMP and EMMPRIN: MCF-7 (300,000 cells/1.5 mL) cells were grown in absence (lane C) and presence of 30 M ATRA (lane E) for 24 hours in SFCM. The cells were collected and were extracted in cell extraction buffer. 50 g of protein from both control and ATRA-treated cell extracts were run on 7.5% SDS-PAGE and the proteins were transferred onto nitrocellulose membrane by western blot. The membrane was incubated with anti MT1-MMP (Figure 3(a)) and EMMPRIN (Figure 3(b)) antibody and then after washing membrane was incubated with respective alkaline phosphatase coupled secondary antibody. Bands were visualized using NBT-BCIP as substrate. Ig-G was used as internal control. The quantitative measurement of the western blot (Figures 3(a) and 3(b)) was performed by Image J Launcher (version 1.4.3.67). (C) is the amount of MT1-MMP and EMMPRIN expression in the control cells, respectively, and (E) is the amount of MT1-MMP and EMMPRIN expression of 30 M ATRA-treated (for 24 hours) MCF-7 cells, respectively.
