Fig. 1a–e

Depletion of ZC3H41 leads to abnormal cell morphologies and defects in cell division. a Effect of ZC3H41 depletion on cell growth. Cells expressing tandem affinity purification (TAP) tag-ZC3H41 were transfected with a plasmid expressing double-stranded RNA (dsRNA) corresponding to ZC3H41 in a tetracycline (tet)-inducible fashion. Cell cultures were followed for up 6 days and diluted every 2 days as needed. Depletion of TAP-ZC3H41 was confirmed by western blot after 2 days of tet induction (inset); RRP4 was included to assess equal loading. b, c Analysis of cell morphology in ZC3H41-depleted cells that were RNA interference (RNAi)-induced for 2 (b) or 3 (c) days with tet. Cells were stained with an anti-EP1 antibody to aid in their visualization, and mounted in mounting medium containing diamidino-2-phenylindole (DAPI). Bar represents 10 µm. A zoid is indicated with an arrow in b; multinucleated cells exhibiting the ‘nozzle phenotype’ are indicated with asterisks in c. d Nucleus (N) and kinetoplast (K) configurations of individual cells were analyzed by DAPI staining. The percentage of cells with physiological (1N1K, 1N2K and 2N2K) or aberrant karyotypes (0N1K, 2N1K and > 2N) is shown for uninduced (− tet) or ZC3H41-depleted cells induced for 1, 2, 3 or 4 days with tet. e Cell cycle analysis of uninduced (− tet) or ZC3H41-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)