Fig. 2a–e

ZC3H41 is found associated with Tb927.7.7460 (Z41AP) in a stable complex. a, b Sodium dodecyl sulfate-polyacrylamide gel electrophoresis/SYPRO staining of TAPs using TAP alone, TAP-ZC3H41 or TAP-Z41AP proteins as baits. The calmodulin binding peptide (CBP) tag remains after tobacco-etch virus (TEV) protease cleavage in the TAP procedure. Proteins identified by mass spectrometry are indicated (a complete list is presented in Additional file 4: Table S2). TAP-ZC3H41 purifications carried out in the absence (−) or presence (+) of RNases A and T1 are shown in b. c Z41AP is essential in procyclic trypanosomes. A cell line expressing TAP-Z41AP was transfected with a plasmid producing dsRNA corresponding to Z41AP in a tet-dependent manner. Cell cultures were followed for up to 6 days and diluted every 2 days as needed. Depletion of TAP-Z41AP was confirmed by immunoblot after 2 days of tet induction (inset; see also e); RRP4 was used to assess equal loading. d Cell cycle analysis of uninduced (− tet) or Z41AP-depleted cells induced for 2 or 3 days with tet. Percentages of cells in sub-G1, G1/S or G2 phases, as well as those having more than two nuclei (> 2N), are expressed as the mean (horizontal lines) ± SEM (shaded areas) of three independent RNAi inductions (dots). e Effect of ZC3H41 or Z41AP depletion on the levels of the corresponding protein partner. A cell line expressing both 4xTy-ZC3H41 and TAP-Z41AP was transfected with plasmids expressing dsRNA corresponding to either ZC3H41 or Z41AP. Protein levels were monitored by immunoblot using BB2 or anti-protein A antisera that detect 4xTy or TAP tags, respectively. α-tubulin was used as a loading control. For other abbreviations, see Fig. 1