Journal: Cell Stress & Chaperones
Article Title: Calnexin overexpression sensitizes recombinant CHO cells to apoptosis induced by sodium butyrate treatment
doi: 10.1007/s12192-008-0054-0
Figure Lengend Snippet: Selection of dox-regulated Tet-TNF-Cnx clones. Western blot analysis of Cnx in the selected overexpressing Tet-TNF-Cnx clones (2, 5, 6, 13, and 24) and a negative control cell (Tet-TNF-Null) to choose highly dox-regulatable clones. Cells were cultivated in the maintenance medium containing 1 μg/ml dox to regulate Cnx expression to a basal level. Two days after seeding, cells were cultivated for 24 h with 2 μg/ml dox or without dox after which they were sampled. Cell lysates were loaded at 5 × 104 cells/10 μl concentration. Relative band intensity was calculated using TINA 2.0 software. Regulation folds were calculated by dividing band intensity of 0 μg/ml dox condition by that of 2 μg/ml dox condition
Article Snippet: Regulation folds were calculated by dividing band intensity of 0 μg/ml dox condition by that of 2 μg/ml dox condition Culture medium and maintenance The medium for culture maintenance of Tet-TNF-Cnx and Tet-TNF-Null cells was IMDM supplemented with 10% dialyzed fetal bovine serum (Gibco, Grand Island, NY, USA), 250 μg/ml hygromycin, 350 μg/ml zeocin, 1 μg/ml dox (Clontech), and 0.02 μM methotrexate (MTX, Sigma).
Techniques: Selection, Clone Assay, Western Blot, Negative Control, Expressing, Concentration Assay, Software