FIGURE 4: Lp uptake by U18666A- and ketoconazole-treated BMDMs. (a) Representative micrographs showing one completely (arrow head) and one partially internalized (arrow) bacterium. BMDMs were infected with GFP+ Lp ΔflaA and stained with anti-Lp antibody prior to permeabilization to distinguish intracellular (green) from surface-associated bacteria (red and green). (b) Phagocytosis of Lp by BMDMs pre-treated for 24 hrs with either DMSO, 3.3 µM ketoconazole or 5 µM U18666A prior to a two hr infection with GFP+ Lp ΔflaA. As a control, some BMDMs were treated with 5 µM cytochalasin D for the duration of the infection. Uptake index is calculated by normalizing the percentage of internalized bacteria for each condition to the percentage of internalized bacteria by the DMSO-treated cells in each experiment. Bars represent average of three biological replicates ± SD where > 100 bacteria for each treatment were scored. (n.s) not-significant; one-way ANOVA. (a-b) Representative data from one of two biological replicates are shown.