JCS -- Wang et al. 120 (2): 320 Figure IG3

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Fig. 3. TNF ${alpha}$ -induced cell death in Cx43-expressing LNCaP cells is greater at higher cell densities. LNCaP cells were plated at different densities, 1250 ( ${circ}$ , $bullet$ ), 2500 ( ${triangleup}$ , ${blacktriangleup}$ ), 5000 ( ${square}$ , ${blacksquare}$ ) or 10,000 ( ${diamond}$ , ${diamondsuit}$ ) initial cells/well. After 12 hours, cells were left uninfected (open symbols) or infected with Ad-Cx43 (filled symbols), incubated for an additional 12 hours and then treated with different concentrations of TNF ${alpha}$ for 48 hours, and cell viability was measured using the MTS assay. At the highest density (10000 cells/well), viability of Ad-Cx43-infected LNCaP cells was significantly reduced by all concentrations of TNF ${alpha}$ (P=0.0015 for 0.01 ng/ml, P<0.0001 for concentrations $>=$ 0.1 ng/ml) as compared to untreated cells. When the initial cell number was decreased to 5000 cells/well, only TNF ${alpha}$ concentrations $>=$ 1 ng/ml induced a significant decrease in cell viability as compared to untreated cells (P<0.0001). At the highest concentrations of TNF ${alpha}$ , slight decreases in cell viability were observed for cells plated at 1250 or 2500 cells per well, but these changes were not statistically significant. In uninfected LNCaP cells, TNF ${alpha}$ only reduced cell viability significantly when used at 10 ng/ml in cells cultured at the highest initial density (viability=85.3±2.2%, P=0.0052).