Journal: The Journal of Biological Chemistry

Article Title: S-nitrosylation of the scaffold protein STRAP enhances oxidative stress–induced apoptosis

doi: 10.1016/j.jbc.2026.111141

Figure Lengend Snippet: Interaction between STRAP and three NOS memebers. A – C , coimmunoprecipitation analysis of STRAP interaction with iNOS ( A ), eNOS ( B ), or nNOS ( C ). HEK293 cells were cotransfected with FLAG-STRAP and iNOS, eNOS, or nNOS expression plasmids. Cell lysates were immunoprecipitated with anti-FLAG antibody or control mouse immunoglobulin G, followed by immunoblotting with anti-iNOS, anti-eNOS, or anti-nNOS antibodies. n = 2. D , detection of S-nitrosylated STRAP (SNO-STRAP) in HEK293 cells overexpressing iNOS, eNOS, or nNOS. Reaction without ascorbate (−Ascorbate) was included as a negative control. n = 3. E , expression levels of iNOS, eNOS, and nNOS in HEK293 cells. eNOS, endothelial nitric oxide synthase; HEK293, human embryonic kidney 293 cell line; iNOS, inducible nitric oxide synthase; NOS, nitric oxide synthase; nNOS, neuronal nitric oxide synthase; SNO, S-nitrosolthiol; STRAP, serine–threonine kinase receptor–associated protein.

Article Snippet: Eluted proteins were separated by SDS-PAGE on 4% to 20% Criterion Precast Midi Protein Gels (Bio-Rad), transferred to PVDF membranes, and probed with the following antibodies: rabbit anti-eNOS (Cell Signaling, 32027S), rabbit anti-nNOS (Cell Signaling, 4231S), rabbit anti-iNOS (Santa Cruz, sc-8310), and rabbit anti-myc (Cell Signaling, 2278S).

Techniques: Expressing, Immunoprecipitation, Control, Western Blot, Negative Control

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

Journal: Neurochemical Research

Article Title: Cerebrolysin Ameliorates Age-Induced Dendritic Spine Degeneration and Memory Decline in C57BL6 Mice

doi: 10.1007/s11064-025-04627-0

Figure Lengend Snippet: Results of immunofluorescence assays. a Immunofluorescence photographs of synaptophysin (red) and the cell nucleus (blue). b Immunofluorescence photographs of Brain Derived Neurotrophic Factor (red) and the cell nucleus (blue). c Immunofluorescence photographs of α-synuclein (red) and the cell nucleus (blue). d Immunofluorescence photographs of neuronal Nitric Oxide Synthase (green) and the cell nucleus (blue). e Immunofluorescence photographs of β-actin (red) and the cell nucleus (blue). f CBL increases synaptophysin in prefrontal cortex and hippocampus. g CBL increases the presence of Brain Derived Neurotrophic Factor in the prefrontal cortex, CA1 hippocampus, and basolateral amygdala. h α-synuclein is not affected by CBL treatment. i CBL treatment did not affect nNOS density. j CBL induces an increase in β-actin in each analyzed region. (PFC: Prefrontal cortex; DG: Dentate gyrus; BLA: basolateral amygdala; Syn; Synaptophysin; BDNF; Brain derived neurotrophic factor; nNOS: neuronal oxide nitric synthase)

Article Snippet: Slides were incubated for 24 h at 4 °C with the primary antibody to Syn (Cell Signaling #36406, Clone: D8F6H, 1:100), BDNF (Abcam, clone: 3C11, 5 μg/mL), α-synuclein (Cell Signaling #2647, clone: Syn204, 1:1000), nNOS (Cell Signaling #4231, Clone: C7D7, 1:200), and β-actin (Cell Signaling #3700, clone: 8H10D10, 1:3000).

Techniques: Immunofluorescence, Derivative Assay