Journal: Science Advances

Article Title: Smaug regulates germ plasm assembly and primordial germ cell number in Drosophila embryos

doi: 10.1126/sciadv.adg7894

Figure Lengend Snippet: ( A ) Box plots (median, 75th and 25th percentiles) showing the number of PGCs in embryos from a variety of wild-type or smg -mutant mothers. P values are from the Kruskal-Wallis one-way analysis of variance (ANOVA) followed by Dunn’s test for multiple comparisons; N values as follows: w 1118 = 79; Df/TM3 = 14; smg 1 /Df = 69; smg 1 / smg 47 = 43; smg 47 /Df = 19; smg 47 / smg 47 = 23. ns, not significant. ( B ) Box plot (median, 75th and 25th percentiles) showing the number of pole buds at NC 9 in embryos from a wild-type or smg -mutant mothers. P value is from the unpaired, one-sided Wilcoxon rank sum test. N values as follows: w 1118 = 19; smg 1 / smg 47 = 18. ( C ) Box plot (median, 75th and 25th percentiles) showing the number of phosphohistone H3 (PH3)–positive PGCs in embryos from a wild-type or smg -mutant mothers. P value is from Bonferroni-corrected unpaired, two-sided Wilcoxon rank sum test. N values as follows: w 1118 = 44; smg 1 /Df = 36; smg 1 / smg 47 = 43. ( D to K ) Confocal images showing 4′,6-diamidino-2-phenylindole (DAPI; blue), VAS (green), and PH3 (red) in wild type ( w 1118 ) (D to G) or in smg mutants (H to K). The smg -mutant genotype of the females from which the embryos shown in (H) to (K) were obtained was smg 1 / Df ( 3L ) Scf-R6 . White arrowheads point to VAS-positive cells that are also PH3 positive. Note that, in smg mutants, the somatic nuclei continue to divide at this stage and, therefore, are PH3-positive but VAS-negative. Three-dimensional reconstructions were used to distinguish the VAS-positive, PH3-positive PGC cells from the VAS-negative, PH3-positive somatic nuclei (see Materials and Methods). Scale bar, 25 μm [applies to (D) to (K)]. See table S2 for data and statistical tests.

Article Snippet: Primary antibodies used for IF were as follows: guinea-pig anti-SMG (1:500) ( ); rabbit anti-GFP (1:500) (Abcam Inc.); rabbit anti-PH3 Ser 10 (1:200) (Upstate USA Inc.); rabbit anti-VAS (1:10,000) ( ); rabbit anti-OSK (1:250) for ( ); rabbit anti-OSK (1:1000) for ( ); rabbit anti-BRU1 (1:500) ( ); and rabbit anti-GCL (1:500) ( ).

Techniques: Mutagenesis

Journal: Stem Cell Reviews and Reports

Article Title: Dbx2 , an Aging-Related Homeobox Gene, Inhibits the Proliferation of Adult Neural Progenitors

doi: 10.1007/s12015-023-10600-7

Figure Lengend Snippet: Constitutive Dbx2 overexpression inhibits the G2/M transition in young adult NSPCs. (A to C) Box-and-whisker plots of the fraction of GFP -NSPCs (blue triangles) and Dbx2 -NSPCs (red triangles) in the G0/G1 (A) , S (B) and G2/M (C) phases of the cell cycle ( n = 4); *, p < 0.05, ***, p < 0.001, Student’s t-test. The + symbols indicate mean percentages. Flow cytometry histograms of PI-stained GFP -NSPC and Dbx2 -NSPC cultures from a representative experiment are shown in (D) and (H) , respectively. (E to G) Box-and-whisker plots of the fraction of Ki67+ (E) , pH3+ total (F) and pH3+ late (G) cells in GFP -NSPC (blue triangles) and Dbx2 -NSPC (red triangles) cultures ( n = 4); *, p < 0.05, **, p < 0.01, Student’s t-test. (I to P) Representative images of GFP -NSPC (I , M , K , O) and Dbx2 -NSPC (J , N , L , P) cultures stained with anti-Ki67 (I , J) or anti-pH3 (K , L) antibodies. Hoechst nuclear staining is shown in (M to P) . Scale bar, 40 µm

Article Snippet: Immunofluorescence analysis with an anti-Ki67 mouse monoclonal antibody (Leica Biosystems ACK02, 1:100) or with an anti-pH3 (Ser 10) rabbit polyclonal antibody (ThermoFisher PA5-17869, 1:100) was performed as previously described [ , ].

Techniques: Over Expression, Whisker Assay, Flow Cytometry, Staining