griffonia simplicifolia lection isolectin b4  (Vector Laboratories)

Journal: Nature Communications

Article Title: Placental TRPV2 supports trophoblast function and normal fetal development

doi: 10.1038/s41467-026-77576-z

Figure Lengend Snippet: a Lectin-cytokeratin double staining at E16.5 showing normal (insert 1) and abnormal (insert 2) labyrinth morphology in Trpv2 −/− placentas. Scale bar = 400 μm and 50 μm. Representative images from n = 12–13 placentas per genotype from 8 independent litters. Similar results were observed in all Trpv2 −/− placentas analysed. b Immunohistochemistry of MCT1 and MCT4, representing differentiated SynTI and SynTII cells, respectively. Arrows indicate SynTI cells with altered morphology. Scale bar = 250 μm and 20 μm. Staining was performed on consecutive sections. Representative images from n = 5–6 placentas per genotype from 6 independent litters with similar results. c Enhanced volcano plot of differentially expressed genes identified by bulk RNA-sequencing of E14.5 labyrinth tissue ( Trpv2 +/+ versus Trpv2 −/− ; 111 downregulated and 56 upregulated genes). Differential expression analysis was performed using DESeq2 with Benjamini-Hochberg correction for multiple testing. n = 6 placentas per genotype (3 male and 3 female) from 5 independent litters. Marker genes for different labyrinth trophoblast populations are indicated. d qRT-PCR validation of selected differentially expressed genes identified by RNA-sequencing. Data are presented as mean values ± SEM. Statistical analysis was performed using two-sided Student’s t tests on ΔCt values. n = 6 placentas per genotype (3 male and 3 female) from 5 independent litters. Exact p-values are also provided in the file. e Cnetplot showing the association between differentially expressed genes and enriched Gene Ontology biological processes. f Overrepresentation analysis showing enrichment of SynTII marker genes among downregulated genes. Statistical significance was determined by hypergeometric testing with correction for multiple testing. g MERscope spatial transcriptomic overview of E16.5 placental sections from wild-type (WT) and Trpv2 -deficient placentas (KO1, KO2). Shown are Bdkrb2 (junctional zone), Pecam1 (fetal endothelial cells), and Vtcn1 (SynTII), illustrating overall placental architecture and organisation of major placental compartments used for subsequent region-based analyses. Scale bar = 1 mm h MERscope spatial transcriptomic overview illustrating region-based annotation of labyrinth tissue used for downstream analyses; wild-type (WT) labyrinth regions and two classes of knockout labyrinth regions: morphologically normal regions (NC) and abnormal regions lacking fetal capillaries (AbC). Scale bar = 1 mm i Heatmap of region-based pseudo-bulk expression profiles derived from MERscope analysis comparing WT, NC and AbC labyrinth regions. Samples and genes are ordered by unsupervised hierarchical clustering. Colour scale indicates relative expression (scaled z-score). j Representative MERscope expression maps of SynTI-associated markers ( Atp2a3, Tgfa and Syna ) and the SynTII precursor-associated marker Slc5a5 in WT and Trpv2 -deficient placentas at E16.5, illustrating the spatial distribution of trophoblast lineage markers in normal and abnormal labyrinth regions. Scale bars = 1 mm.

Article Snippet: Sections were incubated with biotinylated lectin (isolectin B4, B-1205, Vector Laboratories; 1:250, 90 min, 37 °C), which was detected using horseradish peroxide (HRP)-conjugated streptavidin (Agilent P0397; 1:800, 30 min, RT) followed by 3’-Diaminobenzidine (Sigma, 10 min, RT).

Techniques: Double Staining, Morphology, Immunohistochemistry, Staining, Bulk RNA Sequencing, Tissue, Quantitative Proteomics, Analysis, Marker, Quantitative RT-PCR, Biomarker Discovery, RNA Sequencing, Knock-Out, Expressing, Derivative Assay

Journal: Nature Communications

Article Title: Placental TRPV2 supports trophoblast function and normal fetal development

doi: 10.1038/s41467-026-77576-z

Figure Lengend Snippet: a Mendelian inheritance pattern of litters obtained from Gcm1 -Cre wt/tg ; Trpv2 +/- crossed with female Trpv2 F/F mice. Data are shown as litter means ± SEM. b Fetal weight of Trpv2 SynTII-/- and Trpv2 SynTII+/+ fetuses at E16.5, E18.5 and birth. Box plots show the median (centre line); box boundaries indicate the 25th and 75th percentiles; whiskers indicate minimum and maximum values. Statistical analysis was performed using two-way ANOVA with Holm-Šídák’s multiple comparisons test. n = 11 litters at E16.5, n = 15 litters at E18.5 and n = 9 litters at term. c Placental weight of Trpv2 SynTII-/- and Trpv2 SynTII+/+ fetuses at E16.5 and E18.5. Box plots show the median (centre line); box boundaries indicate the 25th and 75th percentiles; whiskers indicate minimum and maximum values. Two-sided unpaired Student’s t test. n = 11 litters at E16.5 and n = 15 litters at E18.5. d Lectin-cytokeratin double staining of E18.5 Trpv2 SynTII-/- placentas showing regions lacking fetal capillaries. Scale bars, 500 μm and 50 μm. Representative images from n = 15 Trpv2 SynTII-/- placentas and n = 10 control placentas collected from 13 independent litters. Similar results were observed in all affected Trpv2 SynTII-/- placentas analysed. e Quantification of fetal capillary-void areas within the labyrinth at E18.5. Data were analysed using a two-sided unpaired Student’s t test. n = 10 control placentas and n = 15 Trpv2 SynTII-/- placentas from 13 independent litters. Exact p -values are also provided in the file.

Article Snippet: Sections were incubated with biotinylated lectin (isolectin B4, B-1205, Vector Laboratories; 1:250, 90 min, 37 °C), which was detected using horseradish peroxide (HRP)-conjugated streptavidin (Agilent P0397; 1:800, 30 min, RT) followed by 3’-Diaminobenzidine (Sigma, 10 min, RT).

Techniques: Analysis, Double Staining, Control

Journal: Investigative Ophthalmology & Visual Science

Article Title: An APE1 Redox Inhibitor Attenuates Pathological Retinal Vascularization by Suppressing FGF2 Angiogenic Signaling

doi: 10.1167/iovs.67.3.47

Figure Lengend Snippet: Refi - 10 reduces pathological neovascularization in oxygen-induced retinopathy (OIR) mice. ( A ) Refi-10 was administered intraperitoneally at P13, and retinas were analyzed at P17. ( B ) Analysis of retinal vasculature. ( a ) Representative IB4-stained whole-mount retinas from normal, OIR, OIR + Refi-10 (25 mg/kg), and OIR + Refi-10 (50 mg/kg) mice at P17. Avascular areas were circled in white . ( b ) Enlarged views of the boxed regions in a . Yellow arrows indicate vascular tufts. ( c ) Neovascularization regions were highlighted in white by ImageJ software. ( d , e ) The quantification of neovascular area and avascular area for each group. Scale bars = 1000 µm. ( C ) Representative H&E staining of retinal sections and quantification of endothelial cell nuclei anterior to the inner limiting membrane. Scale bars = 500 µm. Data are presented as mean ± SD ( n = 6 mice per group; * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001).

Article Snippet: Then, they were incubated with IB4 (1:100, DL-1208-.5; Vector Laboratories, USA) overnight at 4°C.

Techniques: Staining, Software, Membrane