Fig. 4
From: An intrinsic mechanism controls reactivation of neural stem cells by spindle matrix proteins
Chro prevents low-level nuclear Pros to promote and maintain NSC reactivation. a A schematic diagram represents pros regulatory region upstream of translation start site and pros coding sequence region. Primers designed in pros regulatory region (pro1-pro6) and pros CDS region were indicated in blue bars. b Quantification of fold enrichment in ChIP-qPCR assay using anti-Chro-C and control for different pros gene regions. The CDS region of pros is used as control. c Third-instar larval NSCs from control, chro RNAi knockdown and chro − were labelled with Mira, Pros and ToPro-3. Nuclear Pros in NSCs: control, 0%, n = 786; chro RNAi, 74%, n = 385; chro −, 75%, n = 491. d Pros, Mira and Dpn were labelled in the NSCs of various genotypes. Nuclear Pros in NSCs: control, 0%, n = 627; chro 612/Df(3L)ED231, 71%, n = 984; chro 17a/71, 80%, n = 829; chro 8c/71, 73%, n = 405; grh 370, 50%, n = 522; grh 370/Df, 58%, n = 861. e NSC clones from control and Mtor k03905 were labelled with Pros, Dpn and CD8. f Larval brains in chro − and chro − pros 17/+ were labelled by EdU and Dpn. g Quantification of percentages of Dpn+ EdU+ cells in chro − (24 ± 2%) and chro − pros 17/+ (44 ± 2%) larval brains. h Pros, Mira and Dpn were labelled in the NSCs in chro − and chro − pros 17/+. Nuclear Pros in NSCs: chro −, 75%, n = 858; chro − pros 17/+, 57%, n = 1240. i Quantification of percentages of NSCs with cellular extensions in chro − (27 ± 2%) and chro − pros 17/+ (10 ± 1%) larval brains. j, k NSC clones from Mtor k03905 or Mtor k03905 ; pros 17/+ were labelled with Dpn, Mira and CD8 or EdU, Mira and CD8. NSCs with cellular extensions: Mtor k03905, 46%, n = 106; Mtor k03905 ; pros 17/+, 26%, n = 95. EdU− Mira+ NSC clones: Mtor k03905 93%, n = 102; Mtor k03905 ; pros 17/+, 60%, n = 65. All error bars indicate ± SD in b, g and i. *** indicates P < 0.001 in g, i by Student’s t-test. NSC clones were outlined by white dotted lines in e, j and k. White arrows indicated NSCs and arrowheads indicated the cellular extension of NSCs. Scale bars, 10 µm
