FIGURE 2:

Localization of ALBA proteins in stress conditions and colocalization studies. Wild-type procyclic trypanosomes of the Lister 427 strain were subjected to different stresses: either (A) heat shock (42°C for 2 h) or (B) starvation (incubation in PBS for 2 h), fixed with PFA, and analyzed by IFA using the anti-ALBA3 antibody (left, green in merge) together with the anti-DHH1 antibody (middle left, red in merge) and counterstained with DAPI (blue in the phase contrast, right). (C) Cells expressing either ALBA3 or (D) ALBA4 as YFP fusions (left, green in merge) together with the DHH1 protein fused to mCherry (middle left, red in merge) were starved in PBS for 2 h, fixed with PFA. and counterstained with DAPI (blue in phase contrast, right). (E) Cells of the 427 strain starved in PBS for 2 h were analyzed by FISH-RNA to detect the poly(A+) RNA content in the cell (left, green in merge) coupled to IFA using the anti-ALBA3 antibody (middle left, red in merge); right, DNA content (DAPI staining). In E, image acquisition was carried out by confocal microscopy, and one z-plane (200 nm) for all is shown. Scale bar, 5 μm.