Chromatin-associated CENP-A levels decrease during development and are regulated by nuclear import. (a) Representative images of embryos expressing GFP::CENP-A at three different developmental stages and frequency distribution of relative intensity measurements of chromatin GFP::CENP-A during development, from the 2- to 100-cell stage. Relative intensity measurements of chromatin GFP::CENP-A during development are different by one-way ANOVA. n =14 (2 cells), 18 (4 cells), 28 (8 cells), 12 (16 cells), 18 (30–49 cells), 24 (50–69 cells), and 9 (70–100 cells). Error bars represent mean and 95% confidence interval. (b) Representative images of two cell–stage embryos expressing GFP::CENP-A (OD421) after RNAi depletion of control (black circles; n = 72) or rcc-1 RNAi (purple circles; n = 68). Mean GFP::CENP-A relative intensities according to chromosome length are graphed (mean chromosome length measurements at 2-, 4-, 8-, and 16-cell stages taken from Ladouceur et al., 2015. Control conditions are different by one-way ANOVA (P < 0.001), and rcc1 RNAi conditions are not different by one-way ANOVA (P = 0.14). Bars, 5 µm.