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