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: 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

Journal: Clinical and Translational Medicine

Article Title: Niacin promotes motor function recovery after spinal cord injury via Hcar2‐dependent microglia immunometabolic regulation

doi: 10.1002/ctm2.70683

Figure Lengend Snippet: Hcar2 protein expression was upregulated in perilesional microglia following SCI. (A) Schematic timeline of the in vivo experimental design, indicating spinal cord tissue collection points (days post‐injury, dpi) and the definition of the IA and SA regions. (B) Representative Western blot analysis of Hcar2 protein levels in spinal cord lysates at 1, 3, 7 and 14 dpi. β‐Tubulin served as the loading control ( n = 4). (C) Densitometric quantification of Hcar2 protein levels relative to those of β‐Tubulin ( n = 4). (D) Representative confocal images of spinal cord sections at 7 dpi stained for Iba‐1 (microglia, grey) and Hcar2 (green). Nuclei were counterstained with DAPI (blue). Scale bars: 500 µm (overview), 100 µm (inset). (E) Fluorescence intensity line scan profile across a representative Iba‐1 + Hcar2 + cell in the IA region, demonstrating signal colocalization ( n = 5). (F) Quantification of the Hcar2 + area as a percentage of the total Iba‐1 + microglial area ( n = 5). (G) Representative immunofluorescence images of BV2 cells treated with vehicle or LPS (100 ng/mL) for 24 h and stained for Iba‐1 (red) and Hcar2 (green). Scale bar: 100 µm. (H) Quantification of Hcar2 immunofluorescence intensity in BV2 cells ( n = 3 independent experiments). (I) Representative Western blot analysis of Hcar2 protein levels in BV2 cell lysates. (J) Densitometric quantification of Hcar2 protein levels relative to those of β‐tubulin ( n = 3 independent experiments). The data are presented as the means ± SD. Statistical significance was determined via one‐way ANOVA with Tukey's post hoc test. Normal distribution was confirmed using the Shapiro–Wilk test. ** p < .01, *** p < .001 vs. Sham. ### p < .001 vs. SCI‐IA; ns, not significant.

Article Snippet: Murine BV2 microglial cells (PC‐H2025061124; Procell) were propagated in Dulbecco's modified Eagle's medium (DMEM; 11965092; Gibco) enriched with 10% foetal bovine serum (FBS; A5670701; Gibco) and 1% penicillin‐streptomycin ( PB180120 ; Procella) under standard incubation conditions (37°C, 5% CO 2 , humidified atmosphere).

Techniques: Expressing, In Vivo, Western Blot, Control, Staining, Fluorescence, Immunofluorescence

Journal: Clinical and Translational Medicine

Article Title: Niacin promotes motor function recovery after spinal cord injury via Hcar2‐dependent microglia immunometabolic regulation

doi: 10.1002/ctm2.70683

Figure Lengend Snippet: Niacin promoted an anti‐inflammatory phenotype in vitro through reprogramming microglial immunometabolism. (A) Schematic of the in vitro experimental design involving BV2 microglia challenged with LPS (100 ng/mL) and treated with niacin (NA). (B) Representative images of Iba‐1 (green) and Arg‐1 (red) immunofluorescence staining in BV2 cells treated with increasing concentrations of NA (0.1, 0.3 or 1 mM) for 24 h. Scale bar: 100 µm. (C) Quantification of OCR in BV2 cells, showing the NA‐mediated rescue of mitochondrial respiration. (D) Quantification of the red/green fluorescence intensity ratio from JC‐1 staining, which represents the relative mitochondrial membrane potential (ΔΨm). (E) Representative images of JC‐1 staining showing J‐aggregates (red, high potential) and J‐monomers (green, low potential). Scale bar: 100 µm. (F) Quantification of intracellular ATP levels. (G and H) qRT‒PCR analysis of the mRNA expression levels of the proinflammatory cytokines IL‐6 (G) and IL‐1β (H) normalized to that of β‐actin. (I) Schematic of the in vitro experimental design involving shRNA‐mediated Hcar2 knockdown in BV2 cells followed by LPS (100 ng/mL) and NA (0.3 mM) treatment. (J) qRT‒PCR confirmation of Hcar2 mRNA expression indicating knockdown efficiency. (K) Quantification of OCR in Hcar2 ‐knockdown BV2 cells. (L) Quantification of the red/green fluorescence intensity ratio from JC‐1 staining, representing the relative ΔΨm in Hcar2 ‐knockdown cells. (M) Representative images of JC‐1 staining in Hcar2 ‐knockdown BV2 cells showing J‐aggregates (red) and J‐monomers (green). Scale bar: 100 µm. (N) Quantification of intracellular ATP levels in Hcar2 ‐knockdown cells. (O and P) qRT‒PCR analysis of the mRNA expression levels of the proinflammatory cytokines IL‐6 (O) and IL‐1β (P) in Hcar2 ‐knockdown cells, normalized to that of β‐actin. Data are presented as the means ± SD. Statistical significance was determined via one‐way ANOVA with Tukey's post hoc test. Normal distribution was confirmed using the Shapiro–Wilk test. In A‐H, * p < .05, *** p < .001 vs. Control. # p < .05, ## p < .01, ### p < .001 vs. LPS; In I‐P, * p < .05, ** p < .01, *** p < .001 vs. NC+LPS. ### p < .001 vs. NC+LPS+NA; ns, not significant; n = 3 independent experiments.

Article Snippet: Murine BV2 microglial cells (PC‐H2025061124; Procell) were propagated in Dulbecco's modified Eagle's medium (DMEM; 11965092; Gibco) enriched with 10% foetal bovine serum (FBS; A5670701; Gibco) and 1% penicillin‐streptomycin ( PB180120 ; Procella) under standard incubation conditions (37°C, 5% CO 2 , humidified atmosphere).

Techniques: In Vitro, Immunofluorescence, Staining, Fluorescence, Membrane, Expressing, shRNA, Knockdown, Control

Journal: Brain, Behavior, & Immunity - Health

Article Title: Physical activity-associated extracellular vesicles inhibit inflammagen-induced microglial activation

doi: 10.1016/j.bbih.2026.101200

Figure Lengend Snippet: Plasma derived from TMPA rats inhibits the LPS-induced activation of NF-κB in BV2 microglial cells. (A) Timeline for experiments identifying the optimal dosage of LPS treatment for BV2 cell. (B) Representative Western blot images showing phosphorylated levels of molecules involved in MAPK and NF-κB signaling in BV2 cells treated with various LPS doses at indicated time points. (C) Quantitative results of phosphorylated level of p65 in BV2 cells treated with various LPS doses at indicated time points. (D) Quantitative results of phosphorylated level of JNK in BV2 cells treated with various LPS doses at indicated time points. (E) Timeline for experiments evaluating the effects of plasma from SED and TMPA rats on p65 and JNK phosphorylation in BV2 cells treated with 100 ng/mL of LPS. (F) Quantitative results of cell viability using CCK-8 assay. (G) Representative Western blot images showing phosphorylated levels of p65 and JNK in BV2 cells treated with plasma from SED or TMPA rats followed by 100 ng/mL of LPS at indicated time points. (H) Quantitative results of phosphorylated level of p65 in BV2 cells treated with plasma from SED or TMPA rats followed by 100 ng/mL of LPS at indicated time points. (I) Quantitative results of phosphorylated level of JNK in BV2 cells treated with plasma from SED or TMPA rats followed by 100 ng/mL of LPS at indicated time points. Data in panels (C) and (D) were expressed as the median with interquartile range and analyzed with Kruskal-Wallis test followed by Dunn's multiple comparisons test. Data in panels (F) , (G) and (H) were expressed as mean ± standard deviation and analyzed with ordinary two-way ANOVA followed by Tukey's multiple comparisons test. Statistical significances from post hoc multiple comparisons were indicated by asterisks: * p < 0.05, ** p < 0.01, *** p < 0.001. Asterisks in panels (C) and (D) indicate significant differences compared to the 0 ng/mL group. n = 4 cultures/group in assays shown in panels (B – D) and n = 9 cultures/group in assays shown in panels (F – I) .

Article Snippet: Immortalized murine microglial BV2 cells (Cat#: ABC-TC212S, AcceGen Biotechnology, Fairfield, NJ, USA; RRID: CVCL_0182) were cultured in Dulbecco's modified Eagle's medium/nutrient mixture F-12 (DMEM/F12, Cat#: 12500062, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS) with extremely low endotoxin levels (<0.05 EU/ml; Lot#: VP2002200, Cat#: TMS‐013‐BKR, Merck‐Millipore, Burlington, MA, USA) and penicillin-streptomycin (Cat#: 15140122, Thermo Fisher Scientific).

Techniques: Clinical Proteomics, Derivative Assay, Activation Assay, Western Blot, Phospho-proteomics, CCK-8 Assay, Standard Deviation

Journal: Brain, Behavior, & Immunity - Health

Article Title: Physical activity-associated extracellular vesicles inhibit inflammagen-induced microglial activation

doi: 10.1016/j.bbih.2026.101200

Figure Lengend Snippet: Circulating EVs derived from TMPA rats inhibits the LPS-induced activation of NF-κB in BV2 microglial cells. (A) Representative fluorescence images from the EV uptake assay in BV2 cells. The images in the rightmost column are magnified views of the areas outlined by red frames in the color-merged images. Scale bar: 50 μm. (B) Quantitative results of EV uptake assay in BV2 cells. (C) Timeline for experiments evaluating the effects of circulating EVs from SED and TMPA rats on p65 and JNK phosphorylation in BV2 cells treated with 100 ng/mL of LPS. (D) Quantitative results of cell viability using CCK-8 assay. (E) Representative Western blot images showing phosphorylated levels of p65 and JNK in BV2 cells treated with circulating EVs from SED or TMPA rats followed by 100 ng/mL of LPS at indicated time points. (F) Quantitative results of phosphorylated level of p65 in BV2 cells treated with circulating EVs from SED or TMPA rats followed by 100 ng/mL of LPS at indicated time points. (G) Quantitative results of phosphorylated level of JNK in BV2 cells treated with circulating EVs from SED or TMPA rats followed by 100 ng/mL of LPS at indicated time points. Data in panel (B) was expressed as the median with 95% confidence interval. Data in panels (D) , (F) and (G) were expressed as mean ± standard deviation and analyzed with ordinary two-way ANOVA followed by Tukey's multiple comparisons test. Statistical significances from post hoc multiple comparisons were indicated by asterisks: *** p < 0.001. n = 3 cultures/group in assays shown in panels (A) and (B) and n = 9 cultures/group in assays shown in panels (D – G) . (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Immortalized murine microglial BV2 cells (Cat#: ABC-TC212S, AcceGen Biotechnology, Fairfield, NJ, USA; RRID: CVCL_0182) were cultured in Dulbecco's modified Eagle's medium/nutrient mixture F-12 (DMEM/F12, Cat#: 12500062, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS) with extremely low endotoxin levels (<0.05 EU/ml; Lot#: VP2002200, Cat#: TMS‐013‐BKR, Merck‐Millipore, Burlington, MA, USA) and penicillin-streptomycin (Cat#: 15140122, Thermo Fisher Scientific).

Techniques: Derivative Assay, Activation Assay, Fluorescence, Phospho-proteomics, CCK-8 Assay, Western Blot, Standard Deviation