Figure 3

Adherent viable cells isolated from LPI BALF respond to SP-D and GM-CSF and increase uptake of protein and dying cells. (A) LPI BALF contained foamy macrophage cells that did not take up protein. These cells did not survive even an over night culturing, and thus, was not included in the quantification of uptake index. (B) Pre-treatment of primary cells isolated from the BALF with SP-D and GM-CSF facilitate the transport of Alexa-647 conjugated BSA to perinuclear regions. Cells were quantified and presented as means ± SEM of percentage of cells. n = three independent experiments. At least 70 cells were counted for each condition. Uptake index represents the proportion of cells that have Alexa-647 signal in the perinuclear region out of total cell present. The means were compared with respective controls using Dunnett's multiple mean comparison procedure (* p < 0.05). (C) Presence of SP-D and GM-CSF increase the uptake of apoptotic cells by LPI BAL cells. n = three independent experiments. The uptake index represents the proportion of cells that show uptake of apoptotic material out of total cell present. The means were compared with respective controls using Dunnett's multiple mean comparison procedure (* p < 0.05). (D) Representative images showing different categories of cells found 30-min after BSA feeding. (E) Representative images showing the two categories: No uptake; Uptake, where internalized apoptotic bodies/cells were present inside the macrophages.