Figure 6.

Tc38 patterns in T. cruzi epimastigotes synchronized with hydroxyurea. Tc38-Alexa 488 signal is shown in green and DAPI nucleic acid staining in blue. "N" indicates the nucleus and "k" indicates the kinetoplast. (A) Single confocal sections (~0.3 μm thick) of selected parasites that show the most frequent patterns seen in the cell cycle progression after hydroxyurea removal, at the indicated times. Upper panels show the DAPI blue signal, middle panels the Tc38 signal and bottom panels the merge of both. The same patterns were observed in three different synchronization experiments. (B) Z projection of 31 optical sections (~0.3 μm thick) of three selected parasites at 6 h after HU removal. Only the merge of the DAPI and Alexa-488 signals is shown. (C) Western blot of total protein extract using purified anti-Tc38 antibody. M: molecular weight markers, A: protein extracts of asynchronous epimastigote cultures in exponential growth phase. Remaining lanes correspond to protein extracts of epimastigote cultures after removal of HU at the times (hours) indicated above each lane. 1 × 10⁷ cells were loaded onto each lane. Molecular weights of the protein ladder are indicated on the left of the gel (kDa).