Fig. 2.
DRBD2 cellular localization and expression pattern throughout metacyclogenesis. a Detection of TAP-tagged DRBD2 by indirect immunofluorescence. DIC: differential interference contrast; DAPI: DNA stained with 4′,6-diamidino-2-phenylindole (DAPI); TcDRBD2: endogenous TAP-tagged DRBD2 detected by anti-protein A primary antibody (1:40,000 dilution), and secondary antibody conjugated to Alexa 488 (1:400 dilution); Merge: merge between TcDRBD2 and DAPI images. b Detection of DRBD2 throughout metacyclogenesis by immunoblotting. Epi exp. - Epimastigotes in exponential growth; Epi late - Epimastigotes in late exponential growth; Str - Epimastigotes subjected to nutritional stress; Meta - Metacyclic trypomastigotes. Estimated molecular weight of TAP-tagged DRBD2: 52 kDa; TcDHH1 (control): 47 kDa. M: BenchMark™ Protein Ladder (Thermo Fisher Scientific). Bound antibodies were detected by the Odyssey® imager. Approximately 5,0 × 106 parasites per well were used