Fig. 7. T. gondii upregulates protein levels of Mcl-1, but not Bcl-2, in human-derived cell lines. Thirty minutes after infection at a parasite to host ratio of 30:1, HL-60 and U937 cells were treated for 8 hours with 5 µg/ml actinomycin D or 40 ng/ml TNF- in combination with 2 µg/ml cycloheximide, respectively. Antigenic lysates were then separated by SDS-PAGE under reducing conditions, proteins transferred to nitrocellulose membranes and Bcl-2 (A) and Mcl-1 (B) were detected by immunostaining using enhanced chemiluminescence detection. T. gondii-induced alterations of Bcl-2 and Mcl-1 protein levels were quantified densitometrically (C). Bars represent the relative changes in protein levels after parasitic infection of untreated cells (open bars) or those treated with pro-apoptotic stimuli (closed bars) compared with levels in uninfected cells. Horizontal dashed lines indicate an unchanged protein level after parasitic infection compared with levels in uninfected controls.