Figure 1

TcHDAC4 is expressed throughout the T. cruzi life cycle with higher levels present in the insect-equivalent stages of the life cycle reproduced in vitro and its knockout affects cell morphology during metacyclogenesis. (A) Immunoblot using anti- TcHDAC4 serum in cell extracts from epimastigote (Ep), metacyclic trypomastigote (Me), amastigote (Am) and cell culture-derived trypomastigote (Tr) cells. α-TcGAPDH was used as a loading control. Only the part of the immunoblot corresponding to position of the TcHDAC4 and TcGAPDH bands is shown. Complete image of the western blot is presented in Supplementary Figure CI-1. (B) Proliferation curves obtained from biological triplicates. (C) Panoptic staining of epimastigote and metacyclic trypomastigote cells from wild type (WT) and TcHDAC4 null mutant (KO) cells. Scale bars = 5 µm. (D) Scanning electron microscopy of metacyclic trypomastigote from wild type (WT) and TcHDAC4 null mutant (KO) cells. (E) Transmission electron microscopy of metacyclic trypomastigotes from wild type (WT) and TcHDAC4 null mutant (KO) cells. The lower panels correspond to larger magnification of the nuclear region of the same cells shown in the upper panel. n: nucleus, k: kinetoplast, f: flagellum.