Journal: Journal of Heredity

Article Title: Differences in Cell Proliferation and Craniofacial Phenotype of Closely Related Species in the Pupfish Genus Cyprinodon

doi: 10.1093/jhered/esz074

Figure Lengend Snippet: Immunofluorescence light sheet imaging visualizes number and location of mitotic cells as indicated by assay for pH3 in the heads of hatching-age Cyprinodon. (A) Regions of the head analyzed for proliferating cells. (B) 3D reconstructions in lateral view of the head for the DAPI channel (gray, stains nuclei), the pH3 channel (red, dividing cells), and both channels merged. Note that pH3 positive cells (red dots) are mainly localized to ventral structures as shown in lateral view. (C, D) Single 2D images in (C) frontal view and (D) lateral view show pH3 cells (red dots) localizing to epithelial and mesenchyme tissues surrounding cartilage elements as exemplified by the ceratohyal. Cartilage cells are identifiable in the DAPI channel (gray) by their widely spaced nuclei indicative of large cuboidal cells. Cartilage elements can be identified by shape, and are outlined by the brightly labeled perichondrium, a dense layer of mesenchyme cells that surrounds cartilage elements. Inset shows region outlined by white box and arrows point to pH3 positive cells in perichondrium. Note also the clusters of pH3 positive cells in jaws (arrow head), especially around lateral edge of jaws as seen in panel C. Labels: bh, basihyal cartilage; br, brain; ch, ceratohyal cartilage; ey, eye; ga, gill arches; lj, lower jaw; pa, pharynx; pf, pectoral fin; uj, upper jaw.

Article Snippet: Cells were labeled for pH3 with primary rabbit anti-pH3 polyclonal antibody (MyBioSource, MBS2517807), and secondary antibody Alexa Fluor 568 goat anti-Rabbit (Thermo Fisher, A11011).

Techniques: Immunofluorescence, Imaging, Labeling

Journal: Journal of Heredity

Article Title: Differences in Cell Proliferation and Craniofacial Phenotype of Closely Related Species in the Pupfish Genus Cyprinodon

doi: 10.1093/jhered/esz074

Figure Lengend Snippet: Cell proliferation varies among species of Cyprinodon in different regions of the head at hatching. Shown are number of pH3 positive cells relative to either surface area of sampled tissue (A) or volume of sampled tissue (B) for the 3 regions of the head sampled plus the index of relative jaw proliferation (see text). Plotted are values for each sample and boxplots. Samples sizes are snail-eater N = 30, omnivore N = 28, scale-biter N = 19. Significance for all post hoc pairwise comparisons (Tukey) is shown above boxplots and corresponds to Supplementary Table S3. Note that levels of proliferation significantly vary between species when sampling the entire head (e.g., head region standardized to surface area) or regions of the head posterior to the jaws (post-jaw subset), but proliferation does not vary in the jaws (jaw subset). ns, not significant. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: Cells were labeled for pH3 with primary rabbit anti-pH3 polyclonal antibody (MyBioSource, MBS2517807), and secondary antibody Alexa Fluor 568 goat anti-Rabbit (Thermo Fisher, A11011).

Techniques: Sampling

Journal: bioRxiv

Article Title: The Aedes aegypti peritrophic matrix controls arbovirus vector competence through HPx1, a heme–induced peroxidase

doi: 10.1101/2022.06.02.494599

Figure Lengend Snippet: The role of HPx1 in PM assembly. A) Midgut transverse slices at 18 h ABM supplemented with dextran. Green: dextran – FITC. Blue: DAPI nuclear staining. Insets highlight dextran localization. B) Representative images of ROS levels measured by DHE oxidation in individual midguts at 18 h ABM. B’) Quantitative analysis of the fluorescence intensity of oxidized DHE (dsLacZ, n=35; dsHPx1, n=39). C) Quantitative analysis of the fluorescence intensity of oxidized DHE from individual midguts at 18 h ABM (dsLacZ, n=32; dsHPx1 - Ab, n=34; dsHPx1 + Ab, n=27). D) Mitosis index in the mosquito midgut at 18 h ABM (phospho-histone H3) (dsLacZ n=33; dsHPx1 n=38). E) The intestinal microbiota load analyzed through eubacterial ribosomal 16S gene expression by qPCR at 24 h ABM (dsLacZ n=6; dsHPx1 n=7). F) Immune-related gene expression upon HPx1 silencing at 24 h ABM by qPCR (dsLacZ n=8; dsHPx1 n=7). *p<0.05, ****p<0.0001, ns = not significant. Data are the mean +/-SEM. The T test for B’, D, E and F, and one-way ANOVA with Tukey’s posttest for C.

Article Snippet: All samples were incubated overnight with a mouse anti-PH3 primary antibody (1:500, Merck Millipore, Darmstadt - Germany) diluted in blocking solution at 4 °C and then washed 3x for 20 min each in washing solution (PBS, 0.1% Tween 20, 0.25% BSA).

Techniques: Staining, Fluorescence, Gene Expression

Journal: The FASEB Journal

Article Title: Alterations of serine racemase expression determine proliferation and differentiation of neuroblastoma cells

doi: 10.1096/fj.202200394rrr

Figure Lengend Snippet: FIGURE 5 Stable knockdown of SR modulated expression of cell cycle-related proteins. The Srrkd-N2A (sh445) or Srrwt N2A neurons (NC) was cultured in 6-well plates at the density of 400 000 cells/well for 24 h. The cultures were switched into differentiated condition containing 1 mM VPA for 48 h. The cells were harvested for immunoblotting. Knockdown of SR in Srrkd (sh445)-N2A decreased SR, ***p < .0001 (A); increased CyclinD1, **p = .003 (B); decreased P21, ***p < .0001 (C); decreased P27, **p = .003 (D); increased pH3, **p = .005 (E); increased pRB, **p = .0032 (F); increased Ki67, *p = .029 (G), relative to those in Srrwt N2A neurons. Student's t-test was used to compare the differences.

Article Snippet: Rabbit monoclonal anti- pH3 (cat# 53348T, RRID: AB_2799431), rabbit monoclonal anti- CyclinD1 (cat# 55506T), mouse monoclonal anti- P27 (cat# 3698T, RRID: AB_2077 832), and rabbit monoclonal anti- p- Rb (ca# 8516T, RRID: AB_11178658) were from Cell Signaling.

Techniques: Knockdown, Expressing, Cell Culture, Western Blot