FIG 3.

Detection of protein synthesis in P. acanthamoebae during infection of Bax/Bak-deficient HeLa cells. HeLa cells deficient in Bax and Bak were infected with P. acanthamoebae (MOI of 25) for 48 h. Cells were labeled with AHA at time point 47 h p.i. for 1 h, and newly synthesized proteins were detected using a Click-iT reaction. The Click-iT-reaction products are shown in green; blue gives a Hoechst 33342 counterstain to visualize bacterial and host DNA. Some aliquots were treated with cycloheximide (100 ng/ml) and/or rifampin (10 ng/μl) 2 h prior to labeling. Scale bars, 50 μm. Arrows indicate P. acanthamoebae. Pictures are representative of at least three independent experiments.