Journal: Neural Regeneration Research
Article Title: Blood–brain barrier disruption and neuroinflammation in the hippocampus of a cardiac arrest porcine model: Single-cell RNA sequencing analysis
doi: 10.4103/NRR.NRR-D-24-01269
Figure Lengend Snippet: Enhanced intercellular communication drives neuroinflammatory responses following cardiac arrest. (A) Heatmap illustrating the numbers of inferred interactions between different cell types based on LR pairs. (B) Chord diagram of the top 30 LR pairs between cell types. (C, D) Network diagrams of the changes in the number of intercellular communications between the Sham and ROSC6h groups (C) and the Sham and ROSC24h groups (D). Red represents an increase, and blue represents a decrease, in the number of interactions. (E) Visualization of enhanced LR pairs involved in microglia–neutrophil communication after cardiac arrest in the ROSC6h and ROSC24h groups vs. the Sham group. Each dot represents an LR pair, with its position indicating the group. Dot size corresponds to the P value (larger dots indicate lower P values), while the color represents the interaction probability, with the gradient ranging from purple (low probability) to yellow (high probability). (F) Secretion of resistin into the supernatant of the neutrophil–microglia co-culture system over time. (G) Comparison of resistin secretion, with and without OGD/R and the presence of neutrophils. (H) Immunofluorescence analysis and quantification of iNOS expression in microglia. The left panel shows representative images of iNOS (red), IBA1 (green, microglia marker), and DAPI (blue, nuclei), with merged images in the bottom row. The bar graph shows quantification of relative iNOS fluorescence intensity. Statistical significance determined by one-way analysis of variance followed by Tukey’s post hoc test; **** P < 0.0001. Scale bars: 20 μm. IBA1: Ionized calcium binding adaptor molecule 1; iNOS: inducible nitric oxide synthase; LR: ligand receptor; ns: not significant; OGD/R: oxygen-glucose deprivation/reperfusion; ROSC: return of spontaneous circulation.
Article Snippet: They were then incubated with the following primary antibodies at 4°C overnight: rabbit anti-ionized calcium binding adaptor molecule 1 (IBA1) (1:200 dilution; Abcam, Cambridge, UK, Cat# ab178846, RRID: AB_2636859); mouse anti-S100A8 (1:200; Proteintech, Wuhan, China, Cat# 66853-1-Ig, RRID: AB_2882193); rabbit anti-myelin basic protein (MBP) (1:50, Cell Signaling Technology, Danvers, MA, USA; Cat# 78896, RRID: AB_2799920); rabbit anti-myeloperoxidase (MPO) (1:100, Abcam, Cat# ab208670, RRID: AB_2864724); mouse anti-IBA1 (1:200, Santa Cruz Biotechnology, Santa Cruz, CA, USA, Cat# sc-32725, RRID: AB_667733); and rabbit anti-inducible nitric oxide synthase (iNOS) (1:200, Proteintech, Cat# 18985-1-AP, RRID: AB_2782960).
Techniques: Co-Culture Assay, Comparison, Immunofluorescence, Expressing, Marker, Fluorescence, Binding Assay
Bioss is a verified supplier 
Bioss manufactures this product 
