T. cruzi-infected WT and IL-17RA KO mice were given BrdU (1 mg/ml, Sigma-Aldrich)/1% sucrose in the drinking water (carefully protected from light) ad libitum. After 5 days, mice were sacrificed and spleen mononuclear cells were collected and stained with surface antibodies. Incorporated BrdU was detected with a BrdU Flow kit according to the manufacturer's specifications (BD Biosciences).
Apoptosis was determined by Annexin V and 7AAD staining according to the manufacturer's specifications (BD Biosciences). Mitochondrial depolarization (ψ) was measured by FACS using 50 nM TMRE (Invitrogen) as described (24 (link)). When indicated, 7AAD or LIVE/DEAD Fixable Aqua dead cell stain was used in combination to identify live, early apoptotic and late apoptotic/necrotic cells.
Apoptosis was determined by Annexin V and 7AAD staining according to the manufacturer's specifications (BD Biosciences). Mitochondrial depolarization (ψ) was measured by FACS using 50 nM TMRE (Invitrogen) as described (24 (link)). When indicated, 7AAD or LIVE/DEAD Fixable Aqua dead cell stain was used in combination to identify live, early apoptotic and late apoptotic/necrotic cells.
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