Journal: bioRxiv

Article Title: Influenza A virus infection during pregnancy increases transfer of maternal bloodborne molecules to fetal tissues

doi: 10.1101/2025.05.27.656420

Figure Lengend Snippet: ( A ) Representative cross-sections of CLDN5 and CD31 staining in placental samples at E16.5; dotted, hatched, and solid lines delineate the decidua, junctional zone, and labyrinth, respectively. No differences were seen in ( B-D ) CLDN5 or ( F-H ) CD31 MFI/mm 2 across placental layers. ( E ) An increase in Cldn5 gene expression in the whole placenta was observed in only the X31 hi group. One representative fetus per litter was used to generate a litter mean for each group. Data are expressed as means ± SEM; circles represent individual placentas; n per group: Con = 8-10, X31 mod = 6-9, X31 hi = 6-8. Statistical analysis was conducted using a one-way ANOVA with Tukey’s post hoc test; * = p < 0.05. Non-significant p-values indicate main treatment effects (not post hoc). Scale bar = 200 μm.

Article Snippet: Fetal brain antibodies: rabbit anti-CLDN5 (1:1000; ThermoFisher, Catalog no. 34-1600), goat anti-CD31 (1:250; R&D Systems, Catalog no. AF3628SP).

Techniques: Staining, Gene Expression

Journal: bioRxiv

Article Title: Influenza A virus infection during pregnancy increases transfer of maternal bloodborne molecules to fetal tissues

doi: 10.1101/2025.05.27.656420

Figure Lengend Snippet: ( A ) Representative sagittal cross-section of DAPI, CLDN5, and CD31 staining in fetal brain samples at E16.5. ( B ) No significant changes were detected in CLDN5 gene expression in fetal whole brains. ( C ) MFI/mm 2 of CLDN5 at the SVZ was reduced in fetuses exposed to X31 hi compared to the control group. ( D ) No changes were seen in CD31 MFI/mm 2 across groups. Statistical analysis was conducted using a one-way ANOVA with Tukey’s post hoc test. For data containing residuals with unequal variance, Brown-Forsythe and Welch’s ANOVA with Dunnett T3 post hoc multiple comparisons was used. ** = p < 0.01. Data are expressed as means ± SEM; circles represent individual fetuses, with one representative fetus per litter; n per group: Con = 7-10, X31 mod = 7, X31 hi = 7-8. SVZ was delineated (white dots) based on DAPI staining and using reference brain atlases ( ; GENSAT, n.d.; ; ). Scale bar = 200 μm. SVZ = subventricular zone.

Article Snippet: Fetal brain antibodies: rabbit anti-CLDN5 (1:1000; ThermoFisher, Catalog no. 34-1600), goat anti-CD31 (1:250; R&D Systems, Catalog no. AF3628SP).

Techniques: Staining, Gene Expression, Control

Journal: Journal of Cancer

Article Title: Oridonin inhibits tumor angiogenesis and induces vessel normalization in experimental colon cancer

doi: 10.7150/jca.55929

Figure Lengend Snippet: ORI inhibited angiogenesis of colon cancer. (A) Representative images of immunohistochemical analysis of tumor microvessels (×200, scale bar =100 µm). (B) CD31 + MVD was dramatically suppressed in ORI- treated group compared to the Control group. (C) Representative Western blot expressions of VEGF, bFGF, angiostatin and endostatin. (D and E) The levels of VEGF and bFGF were dramatically down-regulated in the ORI-treated group, while (F and G) angiostatin and endostatin were increased by treatment with ORI. * p < 0.05, ** p < 0.01.

Article Snippet: Sections were incubated in rabbit anti-CD31 (1:200, Bioworld), with mouse anti-a-SMA (1:100, Boster) for 1 hour at 37 °C.

Techniques: Immunohistochemical staining, Western Blot

Journal: Journal of Cancer

Article Title: Oridonin inhibits tumor angiogenesis and induces vessel normalization in experimental colon cancer

doi: 10.7150/jca.55929

Figure Lengend Snippet: ORI promoted vessel normalization in colon cancer. (A) Tumor vessels were immunostained for CD31 (green) and pericytes for α-SMA (red). (×400, scale bar =50 µm). (B) The significantly increased perivascular cell coverage in the ORI group was observed from day 5 until day 10 compared to the Control group, the highest of perivascular cell coverage for ORI occurred on day 7.

Article Snippet: Sections were incubated in rabbit anti-CD31 (1:200, Bioworld), with mouse anti-a-SMA (1:100, Boster) for 1 hour at 37 °C.

Techniques:

Journal: Animal Models and Experimental Medicine

Article Title: Bilateral carotid‐jugular arteriovenous graft implantation in an ovine model is safe and durable for facilitation of arteriovenous graft innovation

doi: 10.1002/ame2.70096

Figure Lengend Snippet: Representative imaging of explanted AV grafts. The images include scanning electron microscopy (SEM) in the first column, immunohistochemisty (IHC) in the second column, and hematoxylin and eosin (H&E) staining in the third column. The IHC imaging includes DAPI staining (dark blue) for cell nuclei and CD31 staining (magenta) for endothelial cell junctions, which when observed in the characteristic circular ring‐like staining pattern as seen here, suggests the presence of endothelial cells. Column 3 demonstrates patent sections of graft on the left and occluded sections of the graft on the right. The image on the far‐right side of the figure demonstrates the graft sectioning technique relative to its orientation with “1” corresponding to the venous anastomosis, “2” corresponding to graft mid‐segment, and “3” corresponding to the arterial anastomosis.

Article Snippet: Samples were then incubated with mouse anti‐sheep CD31 antibody (NB100‐65900, Novus Biologicals; 1:50 in 1% BSA/PBS) at 4°C overnight.

Techniques: Imaging, Electron Microscopy, Staining

Journal: JCI insight

Article Title: Purine nucleoside phosphorylase inhibition is an effective approach for the treatment of chemical hemorrhagic cystitis.

doi: 10.1172/jci.insight.176103

Figure Lengend Snippet: Figure 3. 8-AG restores CYP-associated changes in bladder blood flow and proteins linked with oxidative stress and visceral nociception in the rat bladder. (A) Western immunoblotting shows significant alterations by CYP in the expression of nitrotyrosine (n = 12–15 per group), a biomarker for peroxynitrite action in conditions of cell damage and oxidative stress; (B) NLRP3 (n = 8 per group), an intracellular sensor that detects danger signals and results in activation of the NLRP3 inflammasome; (C) TLR4 (n = 12–16 per group), a protein belonging to the pattern recognition receptor family with a key role in amplifying the inflammatory response; (D) HIF-1alpha (n = 7–8 per group), which plays an important role in inflammatory processes; and (E) PECAM-1 (n = 11–13 per group), a cellular adhesion and signaling receptor protein expressed at junctions between endothelial cells. (F) Doppler flow meter measurements revealing a significant increase in bladder blood flow in CYP-rats compared with control untreated rat bladders (n = 12–15 per group). In all cases, treatment with 8-AG blocked CYP-induced changes such that CYP + 8-AG bladders were similar to control, healthy bladders. Upper insets (A–E) show representative bands from Western immunoblotting. Densitometry was normalized to total protein staining (loading control [LC]; representative bands shown in lower inset). Representative bands were run on the same blot but were noncontiguous. Data are presented as means ± SD. Ordinary 1-way ANOVA followed by either Tukey’s post hoc or Newman-Keuls multiple-comparison test was used to evaluate significance; *P < 0.05, **P < 0.01; ***P < 0.0001.

Article Snippet: After proteins were transferred to PVDF membranes, total protein fluorescence of the membrane-bound proteins was imaged on a ChemiDoc MP (Bio-Rad), and the membranes were incubated in 5% (weight/volume; w/v) dried milk dissolved in TBS-T (20 mM Trizma, 137 mM NaCl, 0.1% Tween 20, pH 7.6), rinsed with TBS-T, and incubated overnight at 4°C with primary antibody: cytokeratin 20 (Abcam AB230524), HIF-1alpha (Abcam AB216842), Mst3b (Cell Signaling Technology 4062S), nitrotyrosine (Enzo Life Science BML-SA468-0100), NLRP3 (Abcam AB214185), PECAM (Novus Biologicals NB100-2284), PNPase (Atlas Antibodies HPA001625), TLR4 (Santa Cruz Biotechnology SC-293072), and UPIII (Abcam AB231576) diluted in TBS-T containing 5% (w/v) milk or 5% (w/v BSA).

Techniques: Western Blot, Expressing, Biomarker Discovery, Activation Assay, Control, Staining, Comparison

Journal: APL Bioengineering

Article Title: Geometrically engineered organoid units and their assembly for pre-construction of organ structures

doi: 10.1063/5.0222866

Figure Lengend Snippet: Construction and assembly of human kidney glomerular tissue-like organoid units. (a) Fluorescence-stained frozen sections of tri-cultured spherical organoid units ( n = 3). Each marker protein is stained on three cell types: NHMCs (CD90.1), HUVECs (CD31), and HRGEpCs (Nephrin). Scale bar, 50 μ m. (b) Changes over culture duration in co-localization of marker proteins (mean ± SD, n = 3; 8; or 9 images). (c) Overall view of fluorescence-stained frozen sections of tri-cultured spherical organoid units ( n = 3). Scale bar, 500 μ m. (d) Representative fluorescence-stained images of cellular dynamics around the bonding area between the tri-cultured cubic organoid units ( n = 3, upper row : overall view. Scale bar, 1 mm; lower 5 rows : magnified view. Scale bar, 500 μ m).

Article Snippet: The mouse monoclonal anti-CD31 antibody (Cat# 3528) was purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Fluorescence, Staining, Cell Culture, Marker

Journal: APL Bioengineering

Article Title: Geometrically engineered organoid units and their assembly for pre-construction of organ structures

doi: 10.1063/5.0222866

Figure Lengend Snippet: Assembly of organoid units with different geometries. (a) Overview of how to assemble the organoid units with different geometries. (b) Time-sequenced brightfield images of the assembled MDA-MB-231 organoid units ( n = 3). Scale bar, 1 mm. (c) Live fluorescence images of the assembled MDA-MB-231 organoid units, expressing GFP in the cytoplasm ( n = 3). Upper row : wide-field fluorescence images (raw data), lower row : images processed by the THUNDER imaging system with instant computational clearing (ICC) and extended depth of field (EDF). Scale bars, 1 mm. (d) Representative fluorescence-stained images of the assembled human-kidney glomerular tissue-like organoid units ( n = 3). Each marker protein is stained on three cell types: NHMCs (CD90.1), HUVECs (CD31), and HRGEpCs (Nephrin). Upper 5 rows : wide-field fluorescence images (raw data), lower row : images processed by the THUNDER imaging system with instant computational clearing (ICC) and extended depth of field (EDF). Scale bar, 1 mm.

Article Snippet: The mouse monoclonal anti-CD31 antibody (Cat# 3528) was purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Fluorescence, Expressing, Imaging, Staining, Marker