Figure 3

A-D. Effect of TNF-α on cell viability and expression of p21 and p27 in IκBα mutant LN-18 cells. (A) Wild and mutant IκBα LN-18 cells were treated with TNF-α (10 ng/ml) for 12 hr and cell viability was assessed by MTT assay. The percent cell death was determined considering viability of control cells as 100%. (B) IκBα mutant LN-18 cells were treated with TNF-α (10 ng/ml) for different time points and cleavage of PARP (89 kD fragment) was determined by Western blotting. C) IκBα mutant LN-18 cells grown on coverslips were treated with TNF-α (10 ng/ml) for 3 hr and stained with p21 followed by secondary antibody labeled with Cy3 and DAPI was used for staining nucleus (Magnification 63×). D) Expression of p21 and p27 determined by Western blotting in monolayers (M) and spheroids (S) derived from IκBα mutant LN-18 cells treated with TNF-α for 3 hr. The blots were stripped and reprobed with β-actin, which served as control for equal loading of proteins.