Figure 5: ALKBH4 deficiency interferes with cleavage furrow organization and cell migration.

(a) MRC5 cells were treated with ALKBH4 or CTRL siRNA for 48 h, fixed and immunostained with the indicated antibodies and DAPI. Scale bar, 10 μm. (b) Quantification of cells with mislocalised anillin, NM II, aurora B and Plk1 (shown in a). The histograms represent the mean of three independent experiments (500 cells per condition per experiment). P-values were calculated using a two-tailed t-test. Error bars indicate s.d. (c) Migration assays were performed with a monolayer of confluent Alkbh4^Δ/ΔCre or control (Alkbh4^L/LCre) MEF cells. The migration distances from the edges of the wound were calculated from the time point when the scratch was made (0 h) and 24 h later (illustrated in Supplementary Fig. S4c). Assays were run with primary MEFs derived from two different embryos. P-values were calculated using a two-tailed t-test. Error bars represent s.d. from multiple reading points. (d) MRC5 cells were treated with ALKBH4 or CTRL siRNA for 48 h, fixed and immunostained with the indicated antibodies and DAPI(left). Scale bar, 10 μm. Quantification of multinucleated cells (right). Histogram represents the mean of three independent experiments (500 cells per condition per experiment). P-values were calculated using a two-tailed t-test. Error bars indicate s.d.