Journal: Biomedicines

Article Title: Inhibition of PERK Kinase, an Orchestrator of the Unfolded Protein Response (UPR), Significantly Reduces Apoptosis and Inflammation of Lung Epithelial Cells Triggered by SARS-CoV-2 ORF3a Protein

doi: 10.3390/biomedicines11061585

Figure Lengend Snippet: Transient transfection of A549 cells with ORF3a-mRNA. ( a ) Evaluation of ORF3a mRNA levels at 48, 72 and 96 h post-transfection by real-time PCR. ORF3a mRNA levels were normalized to GAPDH gene. ( b ) Detection of recombinant ORF3a protein levels 48, 72 and 96 h post-transfection by Western blotting. For the detection of recombinant ORF3a, a primary polyclonal anti-His tag antibody was used. ORF3a protein expression normalized to GAPDH protein levels. ( c ) Detection of Green Fluorescent Protein (GFP) protein levels 48, 72 and 96 h post-transfection by Western blotting. For the detection of GFP, a primary polyclonal anti-GFP antibody was used. GFP protein expression normalized to GAPDH protein levels. The notation (***) indicate statistically significant differences ( p ≤ 0.001) compared to control cells.

Article Snippet: PERK polyclonal rabbit primary antibody was obtained from OriGene (OriGene, Rockville, MD, USA).

Techniques: Transfection, Real-time Polymerase Chain Reaction, Recombinant, Western Blot, Expressing, Control

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