Fig. 6.

Severe deficiency of jumu impairs hemocyte division by affecting the cell cycle and cytokinesis. a Quantification of circulating hemocyte numbers in jumu mutant third-instar larvae. b Quantification of circulating hemocyte numbers in third-instar larvae of da > jumu RNAi and Hml > GFP > jumu RNAi (da > ^w1118, da > jumu RNAi, Hml > GFP > ^w1118 and Hml > GFP > jumu RNAi were raised at 25 °C or 29 °C). c-d3 Immunostaining against Tubulin (green) and PH3 (red) shows that the circulating hemocytes that are not undergoing mitosis in jumu double heterozygotes display similar microtubule cytoskeletons to controls (c and d); the circulating hemocytes of controls in metaphase (c1), anaphase (c2), and telophase (c3) display clear spindles and cytokinesis. However, most PH3-positive circulating hemocytes in jumu double heterozygotes show defects in their spindles and cytokinesis (d1 and d2), and no cytokinesis is detectable in cells with divided nuclei (d3). e Quantification of the percentage of PH3-positive cells in third-instar larval circulating hemocytes. f, g PH3 staining (green) of circulating hemocytes isolated from third-instar larvae injected with latex beads (red). h Quantification of phagocytic indexes of latex beads. i-j2 Immunostaining against Tubulin (green) and PH3 (red) shows that jumu knockdown hemocytes display similar mitosis defects to those in jumu double heterozygotes hemocytes. k-o Expression of constitutively active Rho1 (Rho1^V14) or overexpression of Rac1 cannot rescue the enlarged cell size of Hml > jumu RNAi circulating hemocytes (n, o). p Real-time PCR analysis of genes associated with cell cycle and division levels of jumu knockdown hemocytes. For all quantifications: error bars represent the S.E.M; NS, not significant; *P < 0.05; **P < 0.01; ***P < 0.001 (Student’s t-test in a, e, p; one-way ANOVA in b, p). Scale bars: 10 μm (c-d3, i-j2), 20 μm (f, g, k-o)