Journal: PLOS One

Article Title: Impact of blast exposure on visual pathway: Mechanism exploration and novel diagnostic perspectives

doi: 10.1371/journal.pone.0344993

Figure Lengend Snippet: (A) Schematic diagram of the visual pathway and key observation areas in its intracranial segment. (B) Representative images of immunofluorescence staining for NeuN (green) and NLRP3 (red), Iba-1 (grey) in mouse visual cortex slices, Group information is shown in images. The red triangle marks indicate the co-localization of NLRP3 and NeuN. The scale bars for all fluorescence intensity channels and the merge are set at 50 μm, the magnified inset is 20μm. Quantitative analysis of visual cortex immunofluorescence density in Iba-1 (C) and NLRP3 (G) . (H) Fluorescence intensity of NeuN + NLRP3 + / NeuN + , n = 4. (D, E, F, I, J) Western blot analysis of Iba-1, IL-1β, cleaved Gasdermin D and NLRP3 in visual cortex lysates, the molecular mass is indicated in kilodaltons, n = 3. (K-L) Quantitative analysis of representative fluorescence images of NeuN staining with TUNEL labeling in the peri-contusion region from different groups that information is shown in images, Scale bar = 100 μm, n = 4. (M) Quantitative analysis of visual cortex NeuN + cells, n = 4. Error bars indicate mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, ns p > 0.05.

Article Snippet: The following primary antibodies were used: Myelin Basic Protein (MBP) (D8X4Q) XP® Rabbit mAb (1:250, CST, # 78896); βIII Tubulin Mouse monoclonal antibody (1: 300, Abcam, #ab78078), NeuN (E4M5P) Mouse mAb (1:250, CST, # 94403), Iba-1 (E4O4W) XP® Rabbit mAb (1:250, CST, # 17198), and NLRP3 Monoclonal Antibody (1:100, Invitrogen, # MA5−23919).

Techniques: Immunofluorescence, Staining, Fluorescence, Western Blot, TUNEL Assay, Labeling

Journal: PLOS One

Article Title: Impact of blast exposure on visual pathway: Mechanism exploration and novel diagnostic perspectives

doi: 10.1371/journal.pone.0344993

Figure Lengend Snippet: (A) Representative images of immunofluorescence staining for NeuN (green), NLRP3 (red), and Iba-1 (grey) in mouse visual cortex slices, as well as group information, are shown in the images. The red triangle marks indicate the co-localization of NLRP3 and NeuN. The scale bars for all fluorescence intensity channels and the merge are set at 50 μm, the magnified inset is 20 μm, n = 4. (B) Quantitative analysis of visual cortex immunofluorescence density in NLRP3, n = 4. (C) Fluorescence intensity of NeuN + NLRP3 + /NeuN + , n = 4. (D) Quantitative analysis of visual cortex immunofluorescence density in Iba-1, n = 4. (E, F, G, H) Western blot analysis of Iba-1, cleaved Gasdermin D and NLRP3 in visual cortex lysates. The molecular mass is indicated in kilodaltons, n = 3. (I, J) Quantitative analysis of representative fluorescence images of NeuN staining with TUNEL labeling in the peri-contusion region from different groups that information is shown in images, Scale bar = 100 μm, n = 4. (K) Quantitative analysis of visual cortex NeuN+ cells. (L) Representative images of immunofluorescence staining for MBP (green) and βIII-Tubulin (red) in mouse optic nerve slices. Group information is shown in images. Scale bar = 20 μm, n = 4. (M) optic nerve MBP density in different groups, n = 4. (N) optic nerve relative fluorescence intensity of MBP + /β III Tubulin + in different groups, n = 4. (O) TEM analysis of the optic nerve between the BE 24 h and BE 28 d groups, red stars indicate axons with obvious demyelination. Error bars indicate mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, ns p > 0.05.

Article Snippet: The following primary antibodies were used: Myelin Basic Protein (MBP) (D8X4Q) XP® Rabbit mAb (1:250, CST, # 78896); βIII Tubulin Mouse monoclonal antibody (1: 300, Abcam, #ab78078), NeuN (E4M5P) Mouse mAb (1:250, CST, # 94403), Iba-1 (E4O4W) XP® Rabbit mAb (1:250, CST, # 17198), and NLRP3 Monoclonal Antibody (1:100, Invitrogen, # MA5−23919).

Techniques: Immunofluorescence, Staining, Fluorescence, Western Blot, TUNEL Assay, Labeling

Journal: iScience

Article Title: Cerebral organoid exosomes reversed behavioral deficits by repressing NLRP3-mediated neuroinflammation in stress models

doi: 10.1016/j.isci.2026.115069

Figure Lengend Snippet: OExo alleviates LPS-induced microglia activation by suppressing NLRP3 expression (A) Representative images of IBA-1 immunostaining reveal the uptake of PKH26-labeled OExo by IBA-1-positive BV-2 microglia after 48 h of incubation. The white arrows highlight the presence of PKH26-labeled OExo surrounding the nuclei of microglia. (B) Representative images of immunostaining for CD206 (top) and iNOS (bottom) demonstrating BV-2 microglia treated with CON (PBS), OExo (50 μg/mL), LPS (100 ng/mL), and LPS combined with OExo (LPS+OExo) for 24 h. (C and D) Quantitative analysis of the fluorescence intensity of CD206 (C) and iNOS (D). n = 6. (E) q-PCR analysis of the pro-inflammatory cytokines IL-6, IL-1β, and TNF-α in BV-2 microglia treated with CON, OExo, LPS, and LPS+OExo, with glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as the internal reference gene. n = 9. (F) Representative WB images displaying the expression of CD206, Arg-1, and iNOS in BV-2 cells treated with CON, OExo, LPS, and LPS+OExo, with β-actin as the internal control. Full-length blots are presented in . (G) Statistical analysis of CD206, Arg-1, and iNOS expression levels from WB. n = 6. (H) Representative WB images illustrating the NLRP3, Casp-1, and IL-1β expression levels in BV-2 cells treated with CON, OExo, LPS, LPS+OExo, nigericin, and nigericin+LPS+OExo. nigericin, a NLRP3 activator, was utilized to inhibit the rescuing effects of OExo on LPS-induced NLRP3 activation. β-actin was used as the internal control. Full-length blots are presented in . (I) Statistical analysis of NLRP3, Casp-1, and IL-1β expression levels by WB. n = 4. (J) Representative WB images showing NLRP3 levels in BV-2 cells treated with CON, MExo, OExo, LPS, LPS+MExo and LPS+OExo. β-actin was used as the internal control. Full-length blots are presented in . (K) Statistical analysis of NLRP3 expression levels in (J). n = 4. All western blot quantifications are based on at least three biological replicates, with two to three technical replicates per experiment. Scale bars, 50 μm. DAPI was used for nuclear counterstaining in all images. Data are presented as mean ± SEM. Data are subjected to one-way ANOVA followed by the Tukey’s test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: IBA-1 , Protein tech; Wako , Cat# 10904-1-AP; Cat# 019-19741.

Techniques: Activation Assay, Expressing, Immunostaining, Labeling, Incubation, Fluorescence, Control, Western Blot

Journal: iScience

Article Title: Cerebral organoid exosomes reversed behavioral deficits by repressing NLRP3-mediated neuroinflammation in stress models

doi: 10.1016/j.isci.2026.115069

Figure Lengend Snippet: Intranasal administration of OExo attenuated microglia activation in the hippocampus in mouse stress models (A) Representative images of immunofluorescence staining for IBA-1 showing PKH26-labeled OExo co-localized with IBA-1-positive microglia in the hippocampus 24 h after intranasal administration of OExo. Nuclei were counterstained with DAPI. The region within the white dashed box is enlarged in the lower image. White arrows indicate PKH26-labeled exosomes surrounding DAPI-stained nuclei in IBA-1-positive microglia. (B) Low-magnification representative images of IBA-1 and DAPI staining showing the hippocampal subregions CA1, CA3, and DG. (C) Representative immunofluorescence images of IBA-1 in the hippocampal CA1, CA3, and DG subregions. Insets show magnified views of the regions within the dashed boxes. (D) Quantitative analysis of IBA-1-positive microglia in the CA1, CA3, and DG subregions. n = 6. (E) qPCR analysis of pro-inflammatory cytokines ( IL-6 , IL-1β , and TNF-α ) in the hippocampus. GAPDH was used as the internal reference gene. n = 9. (F) Representative western blot images of Arg-1 and iNOS protein expression in the hippocampus. β-actin was used as the loading control. Full-length blots are presented in . (G) Representative WB images of NF-κB, p-NF-κB, NLRP3, IL-1β, and Casp-1 protein expression in the hippocampus. β-actin was used as the loading control. Full-length blots are presented in . (H) Quantitative analysis of Arg-1 and iNOS protein expression from western blot data. n = 6. (I) Quantitative analysis of NF-κB, p-NF-κB, NLRP3, IL-1β, and Casp-1 protein expression from WB data. n = 6. (J) Representative double immunofluorescence staining of NLRP3 and IBA-1 in the hippocampus. The regions within the white dashed boxes are enlarged in the right image. White arrows indicate cells positive for both NLRP3 and IBA-1. (K) Quantitative analysis of cells positive for both NLRP3 and IBA-1 among IBA-1-positive microglia in the hippocampus. n = 6. Scale bars, 20 μm (upper) and 10 μm (bottom) in (A); 1,000 μm in (B); 100 μm in (C); and 20 μm in the magnified insets of (C) and 50 μm in (J). Data are presented as mean ± SEM. The interaction between CMS and OExo was evaluated using two-way ANOVA followed by Tukey’s test (details are shown in ). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: IBA-1 , Protein tech; Wako , Cat# 10904-1-AP; Cat# 019-19741.

Techniques: Activation Assay, Immunofluorescence, Staining, Labeling, Western Blot, Expressing, Control, Double Immunofluorescence Staining

Journal: Cell Reports Medicine

Article Title: Blood-brain barrier-penetrative lipid nanoparticles enable systemic delivery of TRIM11 mRNA to disaggregate Tau in Alzheimer’s disease models

doi: 10.1016/j.xcrm.2026.102685

Figure Lengend Snippet: PLNP-mRNA exhibits enhanced BBB penetration and robust gene transfection in vitro and in vivo (A) Schematic representation of features of PLNP-mRNA. (B) Flow cytometry analysis of cellular uptake of Cy5-labeled PLNP-mRNA in SH-SY5Y cells after 4-h incubation. (C) Corresponding confocal fluorescence images of intracellular Cy5 signal. Scale bars, 10 μm ( n = 3). (D) GFP mRNA transfection efficiency of PLNPs in SH-SY5Y cells after 24-h incubation as determined by flow cytometry and confocal microscopy (inset). Scale bars, 50 μm ( n = 3). (E) In vivo whole-brain fluorescence imaging (left) and (right) quantification of Cy5-labeled PLNP-mRNA distribution in C57BL/6 mice 0.5 h post-intravenous injection (1 mg Cy5-mRNA equiv./kg), showing enhanced brain accumulation relative to controls ( n = 3). (F and G) (F) Ex vivo fluorescence imaging of brains isolated from treated mice and (G) quantification of Cy5 fluorescence intensity specifically in the hippocampus. Scale bars: 1 mm (left) and 300 μm (right) ( n = 3). (H) Representative confocal images showing co-localization of Cy5 signal with neurons (β3-Tubulin), astrocytes (GFAP), and microglia (Iba-1). Scale bars, 50 μm ( n = 3). (I and J) (I) Quantification of Cy5 fluorescence associated with each cell type and (J) fold change in Cy5-positive cell populations ( n = 3 independent experiments). (K) Schematic showing PMPC-mediated targeting of PLNP-mRNA to the BBB via binding to chTs and nAChRs on endothelial cells. (L) Brain-wide GFP expression in C57BL/6 mice 24 h after systemic administration of PLNP-GFP mRNA, visualized by ex vivo fluorescence imaging. Scale bars: 1 mm (left) and 300 μm (right). (M and N) (M) Quantitative analysis of GFP fluorescence and (N) fold-change in GFP-positive cells in the hippocampal region, compared with LNP-mRNA controls ( n = 3). (O) Representative images showing GFP expression and phosphorylated Tau (AT8) in 3×Tg-AD mice following PLNP-mGFP administration. Scale bars, 150 μm. (P) Quantification of GFP fluorescence intensity in treated mice ( n = 3). Data are presented as mean ± SD. For in vitro studies ( B–2D), mRNA concentration was 2 μg/mL. For in vivo experiments ( E–2P), dosage was 1 mg mRNA equiv./kg.

Article Snippet: Iba-1 Polyclonal antibody , Proteintech , Cat#10904-1-AP;RRID: AB_2224377.

Techniques: Transfection, In Vitro, In Vivo, Flow Cytometry, Labeling, Incubation, Fluorescence, Confocal Microscopy, Imaging, Injection, Ex Vivo, Isolation, Binding Assay, Expressing, Concentration Assay

Journal: Cell Reports Medicine

Article Title: Blood-brain barrier-penetrative lipid nanoparticles enable systemic delivery of TRIM11 mRNA to disaggregate Tau in Alzheimer’s disease models

doi: 10.1016/j.xcrm.2026.102685

Figure Lengend Snippet: PLNP-mTRIM11 treatment ameliorates Tau pathology, suppresses neuroinflammation, and protects neuronal integrity in 3×Tg-AD mice (A) Schematic overview of the experimental design for assessing insoluble Tau pathology in the hippocampus following treatment; mRNA dosage: 2 mg equiv/kg per injection. (B) IHC staining of phosphorylated Tau (P-ser396), Tau tangles (AT8), and TRIM11 in hippocampal sections collected 28 days after the final injection of PLNP-mTRIM11 or controls. Scale bars, 50 μm. (C–E) Quantification of TRIM11, P-ser396, and AT8 staining intensities in the hippocampus ( n = 3). (F) Western blot analysis of Tau, TRIM11, phosphorylated Tau (P-ser396), and Tau tangle (AT8) expression in hippocampal lysates ( n = 3). (G) Schematic illustration of TRIM11-mediated reduction in neuroinflammation via cytokine modulation. (H–J) ELISA quantification of (H) anti-inflammatory IL-10, (I) pro-inflammatory TNF-α, and (J) IL-6 levels in hippocampal tissue following treatment ( n = 3). (K–M) (K) Representative IHC images and quantification of (L) astrocytic (GFAP) and (M) microglial (Iba-1) immunoreactivity in the hippocampus. Red arrows indicated activated microglia (Iba-1) and astrocytes (GFAP). Scale bars, 500 μm ( n = 3). (N) Schematic representation of TRIM11’s neuroprotective function through suppression of NFL secretion. (O) Confocal imaging of NFL in hippocampal neurons reveals preserved axonal structure following PLNP-mTRIM11 treatment. Scale bars, 50 μm ( n = 3). Data are presented as mean ± SD. mRNA dosage: 2 mg equiv./kg per injection.

Article Snippet: Iba-1 Polyclonal antibody , Proteintech , Cat#10904-1-AP;RRID: AB_2224377.

Techniques: Injection, Immunohistochemistry, Staining, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Imaging

Journal: Cell Reports Medicine

Article Title: Blood-brain barrier-penetrative lipid nanoparticles enable systemic delivery of TRIM11 mRNA to disaggregate Tau in Alzheimer’s disease models

doi: 10.1016/j.xcrm.2026.102685

Figure Lengend Snippet: PLNP-mTRIM11 treatment prevents early Tau pathology and preserves cognitive and behavioral function in young 3×Tg-AD mice (A) Experimental timeline: 5.5-month-old 3×Tg-AD mice received three intravenous injections of PLNP-mTRIM11 (2 mg mRNA equiv/kg) on days 0, 7, and 14. Behavioral and cognitive assessments were conducted starting on day 47. (B) Total distance traveled in the OFT ( n = 8). (C and D) (C) Discrimination index and (D) PI in the NOR test ( n = 8). (E and F) (E) Total platform crossings ( n = 8) and (F) latency ( n = 6) to platform in the MWM test. (G) Nesting behavior scores ( n = 8), indicating motivation and executive function. (H) Quantitative expression of Tau, TRIM11, phosphorylated Tau (P-ser396), and Tau tangles (AT8) in hippocampal tissue following treatment ( n = 3). (I) Representative immunohistochemical images of hippocampal sections stained for P-Ser396, AT8, GFAP (astrocytes), Iba-1 (microglia; scale bars, 100 μm), and NFL (axons; scale bars, 200 μm). Red arrows indicate positive immunostaining for AT8 (P-Ser202/Thr205), P-Ser396, Iba-1, GFAP, and NFL. (J) Quantification of Iba-1-positive microglia in the hippocampus. ELISA quantification of pro-inflammatory cytokines. (K–M) (K) TNF-α, (L) IL-1β, and (M) IL-6 in brain homogenates ( n = 3). All data are expressed as mean ± SD.

Article Snippet: Iba-1 Polyclonal antibody , Proteintech , Cat#10904-1-AP;RRID: AB_2224377.

Techniques: Expressing, Immunohistochemical staining, Staining, Immunostaining, Enzyme-linked Immunosorbent Assay

Journal: Brain Sciences

Article Title: Cefepime Alleviates Comorbid Pain and Depression Induced by Lipopolysaccharide in Female Mice

doi: 10.3390/brainsci16030306

Figure Lengend Snippet: Effects of CFP on Iba-1 protein levels in the HPC and PFC. ( A ) Effects of CFP (200 mg/kg) on Iba-1 protein levels in the HPC. ( B ) Effects of CFP (200 mg/kg) on Iba-1 protein levels in the PFC. Data are presented as the mean ± SEM ( n = 4); ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: Iba-1 , 1:500 , Santa Cruz , sc-32725.

Techniques: