Journal: Nature Communications

Article Title: Space radiation damage rescued by inhibition of key spaceflight associated miRNAs

doi: 10.1038/s41467-024-48920-y

Figure Lengend Snippet: a Network representation for the DNA DSB repair gene targets for miR-125b-5p, miR-16-5p, and let-7a-5p generated by ClueGO in Cytoscape. As indicated in the figure legend, the color for the edges indicate either the predictions used for the miRNA-mRNA connection or the influence two different nodes will have on each other (i.e. Direct Edges). b Quantification of 53BP1 DNA repair foci in the mature 3D HUVEC microvessel cell culture 1.5 h after irradiation with 0.5 Gy of GCR. Representative images are shown on the right (scale bar = 20 µm). The p-values were determined by two-side multiple pairwise comparison. n = 3 biologically independent samples examined for each conditions and a total of the following random independent field of views for each condition: n = 12 field of views for 0 Gy and n = 19 field of views for both 0.5 Gy and 0.5 Gy + antagomirs. For the boxplot the center line represents the median and the lines extending from both ends of the box indicates the quartile (Q) variability outside Q1 and Q3 to the minimum and maximum values. The schematic of the experiment was created with BioRender.com. DNA DSB pathway-specific Gene Set Enrichment Analysis (GSEA) from c the curated chemical and genetic perturbations and canonical pathways collection (C2) and d the gene ontology (GO) collection using miRNA-sequencing data from different tissues (i.e. liver, heart, soleus muscle, and plasma) from C57BL/6 female mice irradiated with or without 0.5 Gy OF GCR exposure. Mice were euthanized ( N = 10 irradiated and N = 10 sham controls) and tissues were harvested 24 h after irradiation. In Fig. 2 the schematics in panels b and d created with BioRender.com released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license.

Article Snippet: 53BP1 foci were immunostained using a polyclonal antibody against 53BP1 (Catalog #: NB100-304, Novus Biologicals, Littleton, CO, USA) and a counterstain for nuclei with YOYO green (Invitrogen, Carlsbad, CA, USA) followed by Alexa fluor conjugates 495 or 488 (Invitrogen, Carlsbad, CA, USA).

Techniques: Generated, Cell Culture, Irradiation, Comparison, Sequencing

Journal: Bone

Article Title: Abaloparatide promotes bone repair of vertebral defects in ovariectomized rats by increasing bone formation.

doi: 10.1016/j.bone.2024.117056

Figure Lengend Snippet: Fig. 3. ABL increases bone formation without excessive enhancement of bone resorption at the vertebral defect site. (A) Representative images of Hematoxylin- Eosin–stained defected vertebrae after administration. Upper half; overall view, scale bar = 500 μm. Lower half; magnified view, scale bar = 50 μm. (B and C) Representative images of defected vertebrae with immunostaining of ALPase, PHOSPHO1, CatK, and ALPase/PCNA after (B) vehicle or (C) ABL administration. Arrows indicate ALPase/PCNA double-positive preosteoblasts. Scale bar = 50 μm. (D and E) Time-course changes in (D) ALP+Ar/T.Ar and (E) PHOSPHO1+Ar/T.Ar at the vertebral defect site (n = 6 at all timepoints). (F) Time-course changes in N.Oc/BS at the vertebral defect site (n = 6 except for OVX Vehicle With Defect at Week 2 [n = 5]). *P < 0.05, OVX ABL With Defect vs. OVX Vehicle With Defect, †P < 0.05, OVX Vehicle With Defect vs. OVX Vehicle, Tukey’s test.

Article Snippet: Sections were then incubated for 2–3 h at room temperature with rabbit polyclonal antisera against ALPase diluted 1:200 in 1 % BSA-PBS [22], rabbit polyclonal anti-PHOSPHO1 antibody (No. Q8TCT1, Cusabio Technology LLC, Houston, TX, USA) diluted 1:200, and mouse monoclonal anti-CatK antibody (No. F-95, Daiichi Fine Chemical Co., Ltd., Takaoka, Japan) diluted 1:100, followed by incubation with horseradish peroxidase (HRP)-conjugated anti-rabbit IgG (No. ab6112, Abcam plc., Cambridge, Cambridgeshire, UK) diluted 1:100 in 1 % BSA-PBS or HRP-conjugated anti-mouse IgG (No. 61-6520, Chemicon International Inc., Temecula, CA, USA) diluted 1:100.

Techniques: Staining, Immunostaining

Journal: Biomolecules

Article Title: Effect of Topical Programmed Death-Ligand1 on Corneal Epithelium in Dry Eye Mouse.

doi: 10.3390/biom14010068

Figure Lengend Snippet: Figure 3. (A) Immunofluorescence staining showing the expression of (i) PD-L1, (ii) CD4, (iii) CD3e and (iv) IL-17 localized in the corneas in the control, desiccation+PBS and desiccation+PD-L1 group. (B–E) Changes in the percentage of PD-L1, CD4, CD3e and IL-17 positive cells in the cornea. Im- munopositivity for PD-L1 is shown in higher-power fields. Immunopositivity was counted in low-power fields and calculated as relative to the total number of DAPI-positive cells. (F,G) Western blot showing the expression of CD4 and IL-17 in the cornea. (H,I) Relative expression ratio of CD4 and IL-17 to β-actin. In (B–E,H,I), data are presented with bar graph (n = 6, day 10, * p < 0.05; ** p < 0.01; *** p < 0.001 significant difference by one-way ANOVA). Original images of (F,G) can be found in Supplementary Materials.

Article Snippet: The membranes were blocked by incubation with 5% BSA in Trisbuffered saline containing Tween 20 TBST for 1 h at room temperature and incubated overnight at 4 ◦C with specific rabbit polyclonal antibodies against CD4 (1:1000, Catalog No. NBP1-19371; Novus Biologicals, Littleton, CO, USA), IL-17 (1:1000, Catalog No. ab79056; Abcam, Inc.), phosphorylated NF-kB (pNF-kB; 1:1000, Catalog No. 3033S; Cell Signaling Technology, Inc.), NF-kB (1:1000, Catalog No. 51-0500; Invitrogen, Inc.), phosphorylated IkB-α (pIkB-a 1:1000, Catalog No. 9246S; Cell Signaling Technology, Inc.), IkB-α (1:1000, Catalog No. PA5-17888; Invitrogen, Inc.), Bax protein (1:1000, Catalog No. sc-20067; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) and β-actin (1:10,000, Catalog No. 5125S; Cell Signaling Technology, Inc.).

Techniques: Immunofluorescence, Staining, Expressing, Control, Western Blot

Journal: Beilstein Journal of Nanotechnology

Article Title: Study of the reusability and stability of nylon nanofibres as an antibody immobilisation surface

doi: 10.3762/bjnano.15.8

Figure Lengend Snippet: FITC fluorescence of anti-BSA antibody. For each group, the FITC fluorescence data of the immobilised anti-BSA antibody, measured in RFU, are given as percentages relative to the reference group 1, n = 5–6. Stripping treatment with commercial Ag/Ac elution buffer pH 6.6 was performed in all groups except group 1, which was used as the reference in the statistical analysis. One-way ANOVA followed by Newman–Keuls test. Difference from original immunocapture system fluorescence (reference group: group 1): *** p < 0.001.

Article Snippet: FITC-labelled sheep polyclonal antibody against BSA was purchased from Thermo Fisher Scientific Inc.

Techniques: Fluorescence, Stripping Membranes

Journal: Beilstein Journal of Nanotechnology

Article Title: Study of the reusability and stability of nylon nanofibres as an antibody immobilisation surface

doi: 10.3762/bjnano.15.8

Figure Lengend Snippet: FITC fluorescence of anti-BSA antibody. For each group, the FITC fluorescence data of the immobilised anti-BSA antibody, measured in RFU, are given as percentages relative to the reference group 1, n = 5–6. Stripping treatment with ammonium hydroxide buffer pH 11 was performed in all groups except group 1, which was used as the reference in the statistical analysis. One-way ANOVA followed by Newman–Keuls test. Difference from original immunocapture system fluorescence (group 1, reference group): *** p < 0.001.

Article Snippet: FITC-labelled sheep polyclonal antibody against BSA was purchased from Thermo Fisher Scientific Inc.

Techniques: Fluorescence, Stripping Membranes

Journal: Beilstein Journal of Nanotechnology

Article Title: Study of the reusability and stability of nylon nanofibres as an antibody immobilisation surface

doi: 10.3762/bjnano.15.8

Figure Lengend Snippet: FITC fluorescence of anti-BSA antibody. For each group, the FITC fluorescence data of the immobilised anti-BSA antibody, measured in RFU, are given as percentages relative to the reference group 1, n = 5–6. Stripping treatment with glycine buffer pH 2.5 was performed in all groups except group 1, which was used as the reference in the statistical analysis. One-way ANOVA followed by Newman–Keuls test. Difference from original immunocapture system fluorescence (group 1): *** p < 0.001. Difference from bare nanofibers after stripping and reconstructing the immunocapture system (group 6): ΔΔΔ p < 0.001.

Article Snippet: FITC-labelled sheep polyclonal antibody against BSA was purchased from Thermo Fisher Scientific Inc.

Techniques: Fluorescence, Stripping Membranes