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Fig. 3. TNF
-induced cell death in Cx43-expressing LNCaP cells is greater at higher cell densities. LNCaP cells were plated at different densities, 1250 (
, 
), 2500 (
, 
), 5000 (
, 
) or 10,000 (
, 
) 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 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 (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 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, 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 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).
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