Supplementary MaterialsS1 Fig: Glia and trachea endoreplicate in response to pet feeding. right, one EdU-positive and one EdU-negative (white boxes). (I) Quantification of EdU-positive glial cells per brain lobe in response to feeding. (J) Quantity of Repo-positive glial cells per brain hemisphere before and after animal feeding. (K) Single Z image of a brain hemisphere from a knockdown animal. Left and middle panels are color overlays with high magnification of 2 Repo-positive glia shown to the Chloroprocaine HCl right (white boxes). (L) Quantification of glia number in knockdown animals compared to control. (M) Single Z image of a brain hemisphere. Left and middle panels are color overlays of control brain after feeding. A high-magnification image of a tracheal nucleus is usually shown to the right (white boxes). (N) Quantification of EdU in trachea from animals fed standard food or sucrose. (G,I) One-way ANOVA with Tukey post hoc analysis and (F,J,L,N) Student two-tailed test, * 0.05, ** 0.01, *** 0.001. Genotypes of panels outlined in S2 Table and data outlined in S1 Data. RNA interference; EdU, 5-ethynyl-2-deoxyuridine; MB, mushroom body; NB, neuroblast; Repo, reversed polarity.(TIF) pbio.3000721.s001.tif (18M) GUID:?5DF49B3B-F7F0-435C-9822-C55735C8366C S2 Fig: PI3-kinaseCdependent growth regulation in the developing brain. (A) Maximum intensity projection of a single brain hemisphere labeled with EdU and quantification of NB size from indicated genotypes (B). (C,M) Glial and (E,H,I,K) tracheal morphology with quantification of glial surface area (D,N) and tracheal surface area from indicated genotypes (F,J,L). Top panels are colored overlays, with bottom panels showing single-channel grayscale images (A), single channel and mask (C,M), or rendered maximum intensity projection with segmentation of trachea (E,H,I,K). Molecular markers are denoted within panels. (G) Quantification of glial number. (B,D,F,J,L,N) Student two-tailed test, * 0.05, ** 0.01,*** 0.001, error bars, SEM. (G) One-way ANOVA with Tukey post hoc analysis. Genotypes of panels outlined in S2 Table and data outlined in S1 Data. EdU, 5-ethynyl-2-deoxyuridine; NB, neuroblast; PI3-kinase, phosphoinositide 3-kinase.(TIF) pbio.3000721.s002.tif (18M) GUID:?8A9A879E-F39E-418E-9E0E-682529253F78 S3 Fig: Glial identify based on location and GAL4 expression. (A) Single Z picture of a segmented human brain hemisphere with glia subtypes and corresponding GAL4 lines. (B) Final number of glia subtypes before and after pet feeding. Glia type was recognized based on location. Error bars, SEM. Black circles indicate solitary mind hemispheres. (C and E) Remaining panels, coloured overlays Chloroprocaine HCl of a single Z-plane from a mind hemisphere of the indicated genotype, with grayscale images on the right. Molecular markers are denoted within panels. Quantification of glial populations are demonstrated Chloroprocaine HCl in D and F. Red columns depict imply of histoneRFP-expressing glia, and black columns depict imply of glia recognized based on position. Whites boxes indicate RFP-expressing cortex glia IL8RA in (C) and SPG in (E). Genotypes of panels outlined in S2 Table and data outlined in S1 Data. RFP, reddish fluorescent protein; SPG, subperineurial glia.(TIF) pbio.3000721.s003.tif (8.0M) GUID:?1372E763-6A99-468D-9279-029C2CF2FBF9 S4 Fig: Delayed NB reactivation and cortex glial growth in mutants is not due to delays in developmental timing. (A) Package plots of NB diameter of EdU-negative (black) versus EdU-positive (orange) NBs. Column figures indicate quantity of NBs analyzed. (B,C) Quantification of EdU-positive neuroblasts (B) and cortex glial membrane surface (C) area after 48 hours of feeding of indicated genotypes. (D,E) Mouth hook morphology of control and mutants at indicated instances. (ACC) College student two-tailed test, ** 0.01. Genotypes of panels outlined in S2 Table and data outlined in S1 Data. Dilp, insulin-like peptide; EdU, 5-ethynyl-2-deoxyuridine; NB, neuroblast.(TIF) pbio.3000721.s004.tif (6.3M) GUID:?EE29B81D-55B3-48C6-B8A3-B1C5BE6A3EBB S5 Fig: NB EdU incorporation in double mutants. (A,B) EdU-positive NBs after 24 hours of feeding in two times mutants. (A) Maximum intensity projection of a mind hemisphere. Top panel is a coloured overlay (reddish, Dpn; green, EdU), and the bottom panel is definitely a single-channel grayscale image (EdU) below with quantification in (B). Mind hemispheres are defined, and the dotted vertical collection shows the midline. (B) One-way ANOVA with Tukey post hoc analysis. *** 0.001. Genotypes of panels.