On day 6, the immature DCs from each well were harvested, pooled, centrifuged and reseeded at 1×105 cells/mL.
CFSE-labeled, IAV infected mice were harvested 18 hours p.i. Draining mediastinal and peribronchial LN were harvested, pooled and single cell suspensions were prepared.
Cells were harvested, pooled with the culture supernatant, washed once in PBS and fixed in 70% ethanol for cell cycle analysis.
After 6 days, they were harvested, pooled, treated with 20 mM EDTA for 15 minutes, and washed with 1% BSA/PBS in preparation for antibody staining.
Apoptosis was monitored by annexin V binding, measuring the mitochondrial membrane potential and determining the activation of caspase 3. Cells were harvested, pooled with the supernatant, washed once in PBS and processed for the different assays.
Both adherent and nonadherent cells were harvested, pooled, and fixed with 1% paraformaldehyde and 70% ethanol.
Cells were harvested, pooled, and then fixed with 4% paraformaldehyde and 70% ethanol.
The three leaves per plant were then harvested, pooled, and dried in a 65°C oven overnight.
For cell cycle and apoptosis assays, both adherent and non-adherent cells were harvested, pooled and fixed with 90% methanol.
The allantoic fluids were harvested, pooled, centrifuged to remove cellular debris and stored at −70°C until use.
When parasitaemias within a group reached > 10%, blood was harvested, pooled and DNA isolated using the protocol previously described [ 34].
