Article Title: Synergistic effect of therapeutic stem cells expressing cytosine deaminase and interferon-beta via apoptotic pathway in the metastatic mouse model of breast cancer
Figure Lengend Snippet: Alteration of apoptosis and proliferation related protein expression in MDA-MB-231 cells after 5-fluorouracil (5-FU) treatment To confirm the effects of 5-FU, serially diluted 5-FU (0.5, 1.0, and 5.0 μg/ml) and 5-fluorocytosine (5-FC) at 5.0 μg/ml were applied to MDA-MB-231 cells, which were then cultured in 6-well plates. Whole cell lysates were resolved by SDS-PAGE and immunoblotted with specific antibodies to BAX, c-fos, and p21 (Cip1/Waf1). ( A ) Expression of BAX and c-fos protein. ( B ) The relative value of BAX protein levels. (C) The relative value of c-fos protein levels. ( D ) The expression of p21 (Cip1/Waf1) protein. ( E ) Graph of p21 (Cip1/Waf1) protein levels. N: negative control (no treatment with 5-FU or 5-FC), C: 5-FC treatment (5.0 μg/ml), U: 5-FU treatment (0.5, 1.0, and 5.0 μg/ml, from the left). Data are presented as the mean ± SD of three different experiments each performed in triplicate. * p
Article Snippet: Next, the membrane was incubated with primary antibody, mouse monoclonal anti-p21 (Cip1/Waf1) (1:1,000 dilution, Cell Signaling Technology, Inc., Danvers, MA, USA), anti-p53 (1:1,000 dilution, Santa Cruz Biotechnology, Inc. Dallas, TX, USA), anti-BAX (1:1,000 dilution, Cell Signaling Technology, Inc.), anti-GAPDH (1:1,000 dilution, Santa Cruz Biotechnology, Inc.), rabbit polyclonal anti-phospho-Erk1/2 (1:1,000 dilution, Cell Signaling Technology, Inc.), anti-phospho-Akt1/2/3 (1:1,000 dilution, Cell Signaling Technology, Inc.), anti-c-fos (1:2,000 dilution, Abcam plc., Cambridge, UK), or anti-phospho-p38 (1:1,000 dilution, Cell Signaling Technology, Inc.) in 1 × TBS with 3% (w/v) BSA (Sigma-Aldrich Co.) and 0.1% (v/v) Tween 20 overnight at 4°C.
Techniques: Expressing, Multiple Displacement Amplification, Cell Culture, SDS Page, Negative Control