Fig. 3
Colocalization of the TBP and SNAP50 proteins in the T. cruzi nucleus by immunofluorescence microscopy. a Immunodetection of TBP and SNAP50. Indirect immunofluorescence was performed by fixing 6 × 104 permeabilized epimastigotes per slide and incubation with a specific antibody against SNAP50 (dil. 1:100) and Alexa Fluor© 488 goat anti-mouse IgG (H+L) (dil. 1:2000) as secondary antibody (panels 1, 2 and 3). Following incubation with anti-Fab’ fragment antibody (dil. 1:1000), the slides were incubated with anti-TBP antibody (dil. 1:100) and subsequently with Alexa Fluor© 594 goat anti-mouse IgG (H+L) (dil. 1:2000) (panels 4, 5 and 6). Merged images of panels 1 and 4, 2 and 5 and 3 and 6 are shown as panels 7, 8 and 9, respectively. b Immunofluorescence carried out without anti-TBP antibody (blocking control). Homologous slides were incubated with anti-SNAP50 antibody, Alexa Fluor© 488 goat anti-mouse IgG (H+L) and anti-Fab’ fragment antibody (panels 10 and 13) followed by incubation with Alexa Fluor© 594 goat anti-mouse IgG (H+L) (panels 11 and 14). Panels 10–11 and 13–14 as merged images (panels 12 and 15). DAPI staining in a and b for visualization of parasite nuclei (N), nucleoli (n) and kinetoplasts (K). White arrows indicate colocalization of TBP and SNAP50 proteins