Journal: Neural Regeneration Research

Article Title: Physical exercise promotes white matter repair after ischemic stroke

doi: 10.4103/NRR.NRR-D-24-00861

Figure Lengend Snippet: Osteopontin may be the molecular bridge for Treg regulation of microglial function after ischemic stroke. (A) Immunoblotting analyses of OPN 21 days after intervention ( n = 4–5). (B) Experimental design for mRNA extraction from Treg cells. (C) mRNA expression of Spp1 in Treg cells. (D) Experimental design for BV2 microglia in vitro . Created with Microsoft PowerPoint 2019. (E) Expression of inflammation-related genes. (F) More fluorescent microbeads (green) co-localized with microglia (Iba1 + ) (red; Alexa Fluor® 555). Scale bar: 50 μm. Data are expressed as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001 (one-way analysis of variance followed by Tukey’s honestly significant difference test). CM: Conditioned medium; DAPI: 4′,6-diamidino-2-phenylindole; Iba1: ionized calcium-binding adapter molecule 1; IL-10: interleukin 10; iNOS: inducible isoform of nitric oxide synthase; OGD/R: oxygen glucose deprivation/re-oxygenation; OPN: osteopontin; PE: physical exercise; qPCR: quantitative polymerase chain reaction; TGF-β: transforming growth factor-beta; tMCAO: transient middle cerebral artery occlusion; TNF-α: tumor necrosis factor-alpha; Treg cell: regulatory T cell.

Article Snippet: BV2 microglial cells (Procell, Wuhan, China, CL0493, RRID: CVCL_0182) and MO3.13 oligodendrocyte cells (EK-Bioscience, shanghai, China, CC-Y1772, RRID: CVCL_D357) were cultured in Dulbecco’s modified Eagle medium (Gibco) containing 10% fetal bovine serum (ScienCell) at 37°C in a 5% CO 2 incubator.

Techniques: Western Blot, Extraction, Expressing, In Vitro, Binding Assay, Real-time Polymerase Chain Reaction

Journal: Neural Regeneration Research

Article Title: Melatonin alleviates neuroinflammation in ischemic stroke by regulating cyclic GMP-AMP synthase– mediated microglial pyroptosis signaling

doi: 10.4103/NRR.NRR-D-24-01070

Figure Lengend Snippet: Melatonin suppresses NF-κB/NLRP3 inflammasome activation and microglial pyroptosis in the ischemic brain. (A) Representative western blotting images of NF-κB/NLRP3 inflammasome proteins in ischemic brain tissues and sham controls treated with MLT or vehicle at 72 hours after reperfusion after MCAO. (B) Quantification of the western blotting data. n = 6 per group. Data are presented as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001 (one-way analysis of variance followed by Tukey’s post hoc test). (C) Chromatin immunoprecipitation (ChIP) assay of NF-κB and GSDMD in BV2 cells treated with lipopolysaccharide. n = 3 per group. The JASPAR website predicted the existence of a binding site between NF-κB and the GSDMD promoter region ( Additional Figure 1 ). Data are presented as mean ± SD. ** P < 0.01 (Student’s t -test). (D) Double immunostaining for Iba-1 (green) and GSDMD (red) at 72 hours after MCAO in the peri-ischemic area as well as in the corresponding regions of sham control brains, both of which were treated with MLT or vehicle. Nuclei were stained with DAPI (blue). Scale bar: 20 μm (10 μm in the magnified images). Arrowheads indicate GSDMD-double-positive cells with microglia-specific markers. All experiments were conducted in triplicate. cGAS: Cyclic GMP-AMP synthase; DAPI: 4′,6-diamidino-2-phenylindole; GSDMD: gasdermin D; Iba-1: ionized calcium-binding adapter molecule 1; IL: interleukin; MCAO: middle cerebral artery occlusion; MLT: melatonin; NF-κB: nuclear factor kappa-B; NLRP3: NOD-like receptor family pyrin domain-containing protein 3; ns: not significant.

Article Snippet: BV2 cells were obtained from Servicebio (Wuhan, Hubei, China; Cat# STCC20009G-1) and cultivated in Dulbecco’s modified Eagle’s medium (DMEM; Gibco, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS; Gibco), and 100 U/mL penicillin and 100 mg/mL streptomycin (Gibco) at 37°C in a humidified cell incubator.

Techniques: Activation Assay, Western Blot, Chromatin Immunoprecipitation, Binding Assay, Double Immunostaining, Control, Staining

Journal: iScience

Article Title: Propranolol alleviates cerebral infarction through the β2-AR-mediated ERK/NLRP3 pathway

doi: 10.1016/j.isci.2026.115779

Figure Lengend Snippet: Propranolol reduced the expression of NLRP3 and IL-1β in BV2 cells subjected to OGD/R (A) Schematic of cell experiment steps. (B) Western blot analysis of NLRP3 and IL-1β in BV2 cells at 4, 6, 8, 12, and 24 h after OGD/R. (C and D) Quantitative analysis of NLRP3 and IL-1β expression ( n = 3 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 versus the control group. (E) Western blot analysis of NLRP3 and IL-1β in BV2 cells of different treatment groups. (F and G) Quantitative analysis of NLRP3 and IL-1β expression ( n = 3 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 versus the control group; # p < 0.05 and ## p < 0.01 versus the OGD/R group.

Article Snippet: The murine microglial cell line BV2 was obtained from Procell Life Science & Technology Co., Ltd. (Procell, Cat# CL-0493).

Techniques: Expressing, Western Blot, Control

Journal: iScience

Article Title: Propranolol alleviates cerebral infarction through the β2-AR-mediated ERK/NLRP3 pathway

doi: 10.1016/j.isci.2026.115779

Figure Lengend Snippet: Blockade of β2-AR reduced the expression of NLRP3 in BV2 cells subjected to OGD/R (A) Western blot analysis of the effects of dobutamine and betaxolol on NLRP3 and IL-1β expression in BV2 cells. (B and C) Quantitative analysis of NLRP3 and IL-1β expression ( n = 3 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗∗∗ p < 0.001 versus the control group. (D) Western blot analysis of salmeterol and ICI118,551 on NLRP3 and IL-1β expression in BV2 cells. (E and F) Quantitative analysis of NLRP3 and IL-1β expression ( n = 3 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 versus the control group; # p < 0.05 and ## p < 0.01 versus the OGD/R group.

Article Snippet: The murine microglial cell line BV2 was obtained from Procell Life Science & Technology Co., Ltd. (Procell, Cat# CL-0493).

Techniques: Expressing, Western Blot, Control

Journal: iScience

Article Title: Propranolol alleviates cerebral infarction through the β2-AR-mediated ERK/NLRP3 pathway

doi: 10.1016/j.isci.2026.115779

Figure Lengend Snippet: Activation of β2-AR induced the expression of NLRP3 and IL-1β via the ERK1/2 MAPK signaling pathway (A) Western blot analysis of the effects of H-89 and U0126 on NLRP3 and IL-1β expression in BV2 cells. (B and C) Quantitative analysis of NLRP3 and IL-1β expression ( n = 3 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗∗ p < 0.01 and ∗∗∗ p < 0.001 versus the control group; # p < 0.05 versus the OGD/R group. (D) Western blot analysis of p -ERK1/2 expression in BV2 cells. (E) Quantitative analysis of p -ERK1/2 expression ( n = 3 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗∗ p < 0.01 and ∗∗∗ p < 0.001 versus the control group; # p < 0.05 versus the OGD/R group.

Article Snippet: The murine microglial cell line BV2 was obtained from Procell Life Science & Technology Co., Ltd. (Procell, Cat# CL-0493).

Techniques: Activation Assay, Expressing, Western Blot, Control