Fig. 12.

Leukotriene-C4 and Doxorubicin accumulation is decreased in p53-mutant LVCaP cells. A. Leukotriene-C4. Cells were grown at 37°C (LNCaP) or 32°C (LVCaP). 80% confluent LVCaP cells were shifted to 38°C (mt p53) and incubated for 60 hours. Cells were incubated with [³H]-leukotriene-C4 for 2 hr at 38°C, then cooled on ice, washed three times with ice-cold PBS, and dissolved in 0.25 ml 1% SDS. Radioactivity was determined by scintillation counting. a. leukotriene-C4 accumulation in LVCaP and LNCaP cells grown at 38°C. b. leukotriene-C4 accumulation in LVCaP cells grown at 32°C (wt p53) and the same cells grown at 38°C (mt p53). B. Doxorubicin Cells seeded in 24-well plates were grown at 37°C (LNCaP) or 32°C (LVCaP). When cells were 80% confluent, LVCaP cells were shifted to 38°C (mutant p53) and incubated for 60 hours. Cells were then incubated with 2 μM doxorubicin for 2 hr at 37°C (LNCaP) or 38°C (LVCaP). At the end of incubation, cells were washed three times with PBS, then observed under a fluorescence microscope with 100× magnification.