Journal: Molecular Medicine Reports

Article Title: Proteasome inhibitor lactacystin enhances cisplatin cytotoxicity by increasing endoplasmic reticulum stress-associated apoptosis in HeLa cells

doi: 10.3892/mmr.2014.2683

Figure Lengend Snippet: LAC increased cisplatin-induced endoplasmic reticulum stress-associated apoptosis in HeLa human cervical cancer cells. (A) HeLa cells were treated with cisplatin (5 μg/ml) and/or LAC (10 μM) for 12 h. Western blot analysis of grp78, CHOP, caspase-4, cleaved caspase-4, caspase-3 and cleaved caspase-3. (B) Quantitation of the protein levels. Data are presented as the mean ± standard deviation, n=3. * P<0.05 vs. control, # P<0.05 vs. cisplatin. LAC, lactacystin; grp78, glucose-regulated protein-78; CHOP, 153/C/EBP homology protein.

Article Snippet: The membranes were blocked with 5% non-fat dry milk in buffer (10 mM Tris-HCl, pH 7.6; 100 mM NaCl; and 0.1% Tween 20) for 2 h at room temperature and then incubated with the appropriate primary antibodies, including the monoclonal rabbit anti-human Ub and monoclonal rabbit anti-human caspase-3 (1:1,000 dilutions; Epitomics, Burlingame, CA, USA), monoclonal mouse anti-human PDI, monoclonal mouse anti-human p62, monoclonal mouse anti-human Grp78, monoclonal mouse anti-human CHOP, polyclonal rabbit anti-human caspase-4, monoclonal rabbit anti-human caspase-3 and monoclonal mouse anti-human β-actin (1:1,000 dilutions; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) overnight at 4°C, followed by incubation with horseradish peroxidase-conjugated secondary antibody (Hangzhou HuaAn Biotechnology Co.. Ltd., HangZhou, China) at 1:2,000 dilution for 1.5 h at room temperature.

Techniques: Western Blot, Quantitation Assay, Standard Deviation

Journal: Oncology Letters

Article Title: Decreased functional expression of Grp78 and Grp94 inhibits proliferation and attenuates apoptosis in a human gastric cancer cell line in vitro

doi: 10.3892/ol.2014.2831

Figure Lengend Snippet: Immunofluorescence analysis revealing the expression of Grp78 and Grp94 in SGC-7901 cells. (A) Expression of Grp78 susbequent to stable transfection in the (a) experimental, (b) negative control and (c) blank control groups. (B) Expression of Grp94 subsequent to stable transfection in the (a) experimental, (b) negative control and (c) blank control groups. Magnification, ×200.

Article Snippet: After ~8 min, the cells were fixed and made permeable prior to staining with mouse anti-human monoclonal anti-GRP78, anti-GRP94 and anti-talin antibodies (1:50; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA).

Techniques: Immunofluorescence, Expressing, Stable Transfection, Negative Control