rabbit anti-ph3 ser 10  (Upstate Biotechnology Inc)

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

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 research

Article Title: The Forkhead box transcription factor FOXM1 is required for the maintenance of cell proliferation and protection against oxidative stress in human embryonic stem cells.

doi: 10.1016/j.scr.2016.03.007

Figure Lengend Snippet: Fig. 4. FOXM1 knockdown in hESCs impairs cell proliferation with delay at the G2/M phase of the cell cycle. (A) Proliferation of hESCs after transfection with control or FOXM1 siRNA #2 was monitored by NF cell proliferation assay for four days. Relative fold change in cell number was expressed with reference to cell number on day 0. Data are shown as mean ± SEM; *P b 0.05 when compared with control siRNA sample. n = 3. (B) Flow cytometric histograms showing the cell cycle profiles of hESCs transfected with control or FOXM1 siRNA #2 at three days post-transfection. Also shown are the percentages of cells in various cell cycle phases. Data shown are representative of four biological replicates. (C) Quantification of cell cycle distribution revealed an elevated percentage of cells in G2/M phase. Data are shown as mean ± SEM. *P b 0.05 when compared with control siRNA sample. n = 4. (D–F) Immunofluorescence analysis of the mitotic marker phospho-histone H3 Ser10 (PH3) (green) in hESCs treated with control or FOXM1 siRNA at three days post-transfection. DNA was stained with Hoechst. Scale bar = 100 μm. (E) Quantification of PH3-positive cells and mitotic cells in hESCs treated with control or FOXM1 siRNA at three days post-transfection. Mitotic cells were regarded as in prophase, prometaphase, or metaphase/anaphase/telophase depending on the chromosome staining pattern. Data are shown as mean ± SEM. **P b 0.01 when compared to control group. n = 4. (F) Images at higher magnification. FOXM1 siRNA #2-treated cells displayed defects in mitosis and chromosome alignment at the equator (enlarged in inset).

Article Snippet: After treatment with mouse anti-FOXM1 (ab55006; 1:100; Abcam), rabbit anti-OCT4 (H-134; 1:100; Santa Cruz), rabbit anti-phospho-histone H3 Ser 10 (PH3) (06–570; 1:500; Upstate) or mouse anti-SSEA-1 (MAB4301; 1:128; Millipore), cells were incubated with goat antirabbit IgG Alexa Fluor ® 488, goat anti-rabbit IgG Alexa Fluor ® 568, goat anti-mouse IgG Alexa Fluor ® 488 or goat anti-mouse IgG Alexa Fluor ® 568 (Invitrogen).

Techniques: Knockdown, Transfection, Control, Proliferation Assay, Marker, Staining