Figure 4(A) Larval brains of control and cdk1 RNAi (BDSC#35350, #36117, and #40950) under insc­-Gal4 at 24h ALH were stained for Deadpan (Dpn) and Miranda (Mira). Data information: White arrows point to NSCs and yellow arrowheads point to cellular extensions of NSCs (A). Scale bars, 5 μm (A), Control: insc­-Gal4/+ UAS-dicer2 (B) Quantifications of cellular extensions for genotypes in (A). Control: n=942, 11 BL; BDSC#35350: n=1056, 13 BL; BDSC#36117: 16.0%±3.5% (n=1059, 12 BL); BDSC#40950: 16.7%±3.1% (n=933, 11 BL). Data information: Data were presented in quantification graphs as mean ± SD (B, n numbers in (B, were total NSCs counted with total brain lobes measured, while each brain lobe was considered as one biological replicate. Statistical analyses were done comparing among different genotypes using ordinary one-way ANOVA (B, ***P < 0.001. Control: insc­-Gal4/+ UAS-dicer2 (C) Larval brains of control and cdk1 RNAi (BDSC#35350, #36117 and #40950) under insc­-Gal4 at 24h ALH were labelled with Dpn and EdU. Data information: Scale bars, 30 μm (C, Control: insc­-Gal4/+ UAS-dicer2 (D) Quantifications of NSCs without EdU incorporation for genotypes in (C). Control: n=849, 10 BL; BDSC#35350: n=741, 10 BL; BDSC#36117: 87.4%±8.0% (n=697, 10 BL); BDSC#40950: 92.6%±2.4% (n=621, 8 BL). Data information: Data were presented in quantification graphs as mean ± SD n numbers in were total NSCs counted with total brain lobes measured, while each brain lobe was considered as one biological replicate. Statistical analyses were done comparing among different genotypes using ordinary one-way ANOVA D, ***P < 0.001. Control: insc­-Gal4/+ UAS-dicer2 (E) Larval brains of control, UAS-cdk1-Myc, and UASp- cdk1.T14A.Y15F.VFP (cdk1.CA) under insc­-Gal4 at 6h ALH were labelled with Dpn and EdU. Data information: Scale bars, 30 μm Control: insc­-Gal4/+ UAS-dicer2 Quantifications of EdU incorporation (F) In (F), n=345, 6 BL for control; n=524, 8 BL for cdk1-Myc and n=280, 6 BL for cdk1.CA. Data information: Data were presented in quantification graphs as mean ± SD F, n numbers in F, were total NSCs counted with total brain lobes measured, while each brain lobe was considered as one biological replicate. Statistical analyses were done comparing among different genotypes using ordinary one-way ANOVA , F, ***P < 0.001. Control: insc­-Gal4/+ UAS-dicer2 (H) Larval brains of control (ctrl; yw), cdk1-Myc overexpression under insc­-Gal4, pr-set7716/Df(3R) ED5644 (716/Df), and pr-set7716/Df with cdk1-Myc induced with insc-Gal4 at 24h ALH were labelled with Dpn and EdU. (I, J) Quantifications of EdU incorporation (I) and NSC diameters (J) for genotypes in (H). In (I), n=1334, 16 BL for control; n=1299, 15 BL for insc>cdk1; n=1221, 18 BL for pr-set7716/Df and n=1372, 16 BL for pr-set7716/Df with cdk1-Mycinduced with insc-Gal4. In (J), n=153 for control; n=152 for insc>cdk1; n=156 for pr-set7716/Df and n=159 for pr-set7716/Df with cdk1-Myc induced with insc-Gal4.Data information: Data were presented in quantification graphs as mean ± SD I, J). n numbers in I) were total NSCs counted with total brain lobes measured, while each brain lobe was considered as one biological replicate. Statistical analyses were done comparing among different genotypes using ordinary one-way ANOVA ***P < 0.001. Scale bars, 30 μm H). Control: insc­-Gal4/+ UAS-dicer2