Journal: Cell reports

Article Title: Astrocytes Amplify Neuronal Dendritic Volume Transmission Stimulated by Norepinephrine

doi: 10.1016/j.celrep.2019.11.092

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: rabbit anti GFAP , Phosphosolution , Cat# 620-GFAP; RRID:AB_2492124.

Techniques: Virus, Plasmid Preparation, Recombinant, Software

Journal: Biomolecules & therapeutics

Article Title: PEGylated Erythropoietin Protects against Brain Injury in the MCAO-Induced Stroke Model by Blocking NF-κB Activation.

doi: 10.4062/biomolther.2019.147

Figure Lengend Snippet: Fig. 2. Effect of P-EPO on MDA and GSH/GSSG levels and expression of GFAP and Iba-1. Vehicle, 5000 U/kg of P-EPO, was adminis- tered via i.v. before MCAO. Ischemic brain tissues were processed for immunohistochemistry 24 h after reperfusion following MCAO. (A) MDA levels in the brain infarction site was measured using a TBARS assay kit. (B) GSH/GSSG levels in the brain infarction site were mea- sured using a GSH/GSSG ratio detection assay kit. Representative photomicrographs of (C) GFAP-positive astrocytes, (D) Iba-1-positive microglia/macrophages. Each value is presented as mean ± SD from five mice. **p<0.01 significant difference from MCAO mice. ##p<0.01, ###p<0.0001 significant difference from Sham mice (Sham n=6-7, MCAO n=8-9, M-EPO n=10-12).

Article Snippet: Membranes were incubated at room temperature for 2 h with the following specific antibodies: anti-COX-2, anti-IκB, anti-p-IκB, anti- STAT1, anti-p-STAT1, anti- STAT3, anti-pSTAT3, anti- STAT5, anti-p-STAT5 (Cell Signaling Technology, Inc., Beverly, MA, USA), anti-inducible nitric oxide synthase (iNOS) and anti-Glial fibrillary acidic protein (GFAP) (1:1,000 Novus Biologicals, Inc., Littleton, CO, USA), anti-p50, antip65, anti-BAX, anti-cleaved caspase-3, histone H1 (1:1,000; Santa Cruz Biotechnologies, Inc., Santa Cruz, CA, USA), antihEPO (1:1,000, R&D systems, Inc., Minneapolis, MN), antiPEG-2-128, anti-PEG-B47 (1:1,000; Abcam, Cambridge, UK) and anti-β-actin (1:2,500; Sigma, St Louis, MO).

Techniques: Expressing, Immunohistochemistry, TBARS Assay, Detection Assay

Journal: Biomolecules & therapeutics

Article Title: PEGylated Erythropoietin Protects against Brain Injury in the MCAO-Induced Stroke Model by Blocking NF-κB Activation.

doi: 10.4062/biomolther.2019.147

Figure Lengend Snippet: Fig. 3. Effects of P-EPO on inflammatory protein and NF-κB, STAT5 activity. 10 h after reperfusion following 2-h MCAO, vehicle (Sham), 5,000 U/kg of P-EPO was administered via i.v. The ischemic cortex in the striatum was collected for Western blot or immunohistochemistry 24 h after reperfusion following MCAO. (A) Representative Western blots of iNOS, COX-2, GFAP and Iba-1 in the ischemic brain. (B) Rep- resentative Western blots of transcription factors STAT1, STAT3, and STAT5 and phosphorylated form and NF-κB factor p50 and p65. Equal loading was confirmed by monitoring the β-actin and histone-H1 protein levels. (C) The expression levels were quantified by stereological analysis using the ImageJ program. (D) Serum levels of cytokines after MCAO measured by ELISA. Anti-inflammatory cytokines (TNF-α, IL-6, IL-1β) were examined. The data are the mean ± SD each sample was probed in triplicate. n=5/group; *p<0.05, **p<0.01, ***p<0.0001 significant difference from MCAO mice. ##p<0.01, ###p<0.0001 significant difference from Sham mice (Sham n=8-9, MCAO n=8-9, M-EPO n=10-12).

Article Snippet: Membranes were incubated at room temperature for 2 h with the following specific antibodies: anti-COX-2, anti-IκB, anti-p-IκB, anti- STAT1, anti-p-STAT1, anti- STAT3, anti-pSTAT3, anti- STAT5, anti-p-STAT5 (Cell Signaling Technology, Inc., Beverly, MA, USA), anti-inducible nitric oxide synthase (iNOS) and anti-Glial fibrillary acidic protein (GFAP) (1:1,000 Novus Biologicals, Inc., Littleton, CO, USA), anti-p50, antip65, anti-BAX, anti-cleaved caspase-3, histone H1 (1:1,000; Santa Cruz Biotechnologies, Inc., Santa Cruz, CA, USA), antihEPO (1:1,000, R&D systems, Inc., Minneapolis, MN), antiPEG-2-128, anti-PEG-B47 (1:1,000; Abcam, Cambridge, UK) and anti-β-actin (1:2,500; Sigma, St Louis, MO).

Techniques: Activity Assay, Western Blot, Immunohistochemistry, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Cervical spinal cord stimulation exerts anti-epileptic effects in a rat model of epileptic seizure through the suppression of CCL2-mediated cascades

doi: 10.1038/s41598-024-64972-y

Figure Lengend Snippet: ( A , C ) Double immunofluorescence staining of DAPI (blue) with Iba-1 (green) and GFAP (red) in the hippocampal CA1, CA3, and DG 7 days after the administration of KA. Scale bar = 80 µm. ( B , D ) Quantitative data of d showing the number of Iba1- and GFAP-positive cells in the hippocampus. Data are presented as the mean ± standard error. n = 7 rats for each group. * p < 0.05, ** p < 0.01, *** p < 0.001. Abbreviations: Iba-1, ionized calcium-binding adapter molecule 1; GFAP, glial fibrillary acidic protein; DAPI, 4,6-diamidino-2-phenylindole; DG, dentate gyrus; KA, kainic acid.

Article Snippet: The following primary antibodies were used for tissue staining: rabbit anti-Iba1 antibody (1:250; Wako Pure Chemical Industries, Osaka, Japan), rabbit anti-GFAP antibody (1:1000; Novus Biologicals, Littleton, CO).

Techniques: Double Immunofluorescence Staining, Binding Assay

Journal: International Journal of Molecular Sciences

Article Title: Cold Atmospheric Plasma Does Not Affect Stellate Cells Phenotype in Pancreatic Cancer Tissue in Ovo

doi: 10.3390/ijms23041954

Figure Lengend Snippet: CAP does not alter the activation profile of RLT-PSC cells in ovo. Sections of RLT-PSC and co-cultured RLT-PSC + Mia PaCa-2 from in ovo tissue were stained for GFAP, vimentin (Vim), and ACTA-2. ( a – c , g – i ) Mean fluorescence intensity (MFI) expressed as arbitrary units (a.u.); ( d – f , j – l ) number of cells positive for the corresponding markers. Anti-CD44 antibodies (PDAC cells) were used as a counterstain to discriminate Mia PaCa-2 cells from RLT-PSC. UT = untreated control. n ≥ 4 tissue per condition; each dot represents one tissue. Dashed lines = median; dotted lines = 25% and 75% quartiles; * = p ≤ 0.05.

Article Snippet: Primary antibodies used: GFAP (HPA056030, Atlas Antibodies, 1/200, Bromma, Switzerland); ACTA-2 (M0851, Dako, 1/200); Vimentin (Abcam ab92547, 1/500, Cambridge, UK); CD44 (3570S, Cell Signaling Technologies, 1/400, Danvers, MA, USA); MMP2 (40994, Cell Signaling Technologies, 1/400, Danvers, MA, USA); MMP9 (13667, Cell Signaling Technologies, 1/100, Danvers, MA, USA).

Techniques: Activation Assay, In Ovo, Cell Culture, Staining, Fluorescence, Control

Journal: International Journal of Molecular Sciences

Article Title: Cold Atmospheric Plasma Does Not Affect Stellate Cells Phenotype in Pancreatic Cancer Tissue in Ovo

doi: 10.3390/ijms23041954

Figure Lengend Snippet: List of primers used for quantitative real-time PCR.

Article Snippet: Primary antibodies used: GFAP (HPA056030, Atlas Antibodies, 1/200, Bromma, Switzerland); ACTA-2 (M0851, Dako, 1/200); Vimentin (Abcam ab92547, 1/500, Cambridge, UK); CD44 (3570S, Cell Signaling Technologies, 1/400, Danvers, MA, USA); MMP2 (40994, Cell Signaling Technologies, 1/400, Danvers, MA, USA); MMP9 (13667, Cell Signaling Technologies, 1/100, Danvers, MA, USA).

Techniques: