Journal: Experimental and therapeutic medicine

Article Title: Study of dietary‑induced progression of psoriasis‑like mice based on gut macrophage polarization.

doi: 10.3892/etm.2023.11976

Figure Lengend Snippet: Figure 5. Protein expressions of Jagged1/Notch1/HES5 and TLR‑2/IκB‑α/NF‑κB p65 signaling pathway on skin lesions of imiquimod‑induced psoriasis model. (A and a) Western blot analysis of Notch signaling and NRP1 in skin lesions. The mammalian Notch signaling pathway consists of four Notch receptors (Notch 1, 2, 3, and 4) and five Notch ligands (Jagged 1, and 2, and Delta‑like 1, 3 and 4). HES5 is an important downstream target gene of Notch signaling pathway. NRP1 is involved in keratinocyte hyperproliferation of psoriasis accompanied by inflammation activated angiogenesis. (B and b) Western blot analysis of TLR‑2/IκB‑α/NF‑κB p65 signaling in skin lesions. Values are expressed as mean ± SD (*P<0.05, **P<0.01 vs. the normal control; #P<0.05, ##P<0.01, ###P<0.001 vs. the psoriasis group) and were statistically analyzed with one‑way ANOVA. NC, normal control; P, psoriasis; P + SF, psoriasis + SF; SF, stimulating food; p‑, phosphorylated.

Article Snippet: Proteins were transferred onto a PVDF membrane (MilliporeSigma), and then incubated with primary antibodies specific for NF‐κB p65 (cat. no. 6956), phospho‐NF‐κB p65 (cat. no. 3033), Toll‐like receptor‐2 (cat. no. 2229S), IKBα (cat. no. 9242), Jagged‐1 (cat. no. 70109s), Notch‐1 (cat. no. 3608s; all CST; 1:1,000), Hes‐5 (cat. no. ab25374; Abcam; 1:1,000) and anti‐β‐actin (cat. no. BM0627; Wuhan Boster Biological Technology, Ltd.; 1:2,000) overnight at 4 ̊C and then followed by incubation with horseradish peroxidase‐conjugated secondary anti‐mouse or anti‐rabbit IgG (cat. nos.

Techniques: Western Blot, Control

Journal: Molecular Medicine Reports

Article Title: Astragaloside attenuates myocardial injury in a rat model of acute myocardial infarction by upregulating hypoxia inducible factor-1α and Notch1/Jagged1 signaling

doi: 10.3892/mmr.2017.6522

Figure Lengend Snippet: Effects of Astragaloside treatment on HIF-1α, Notch1 and Jagged1 mRNA expression levels. (A) HIF-1α, (B) Notch1 and (C) Jagged1 mRNA expression levels were determined by reverse transcription-quantitative polymerase chain reaction. Data are expressed as the mean ± standard deviation (n=6). MI, myocardial infarction; AST, Astragaloside; HIF-1α, hypoxia-inducible factor 1-α.

Article Snippet: Polyclonal anti-HIF-1α (cat. no. #4426) and monoclonal anti-Notch1 (cat. no. #3608) primary antibodies were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA), and monoclonal anti-Jagged1 (cat. no. ab124524) and anti-GAPDH (cat. no. ab181602) antibodies were purchased from Abcam (Cambridge, UK).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: Astragaloside attenuates myocardial injury in a rat model of acute myocardial infarction by upregulating hypoxia inducible factor-1α and Notch1/Jagged1 signaling

doi: 10.3892/mmr.2017.6522

Figure Lengend Snippet: Effects of Astragaloside treatment on HIF-1α, Notch1 and Jagged1 protein expression levels. Representative western blotting images and quantification of (A) HIF-1α, (B) Notch1 and (C) Jagged1 protein expression levels. GAPDH served as an internal control. Data are expressed as the mean ± standard deviation (n=6). 1, sham control group; 2, myocardial infarction model group; 3, 2.5 mg/kg/day Astragaloside treatment group; 4, 10 mg/kg/day Astragaloside treatment group; MI, myocardial infarction; AST, Astragaloside; HIF-1α, hypoxia-inducible factor 1-α.

Article Snippet: Polyclonal anti-HIF-1α (cat. no. #4426) and monoclonal anti-Notch1 (cat. no. #3608) primary antibodies were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA), and monoclonal anti-Jagged1 (cat. no. ab124524) and anti-GAPDH (cat. no. ab181602) antibodies were purchased from Abcam (Cambridge, UK).

Techniques: Expressing, Western Blot, Standard Deviation

Journal: PLoS ONE

Article Title: Neurogenin 3 Expressing Cells in the Human Exocrine Pancreas Have the Capacity for Endocrine Cell Fate

doi: 10.1371/journal.pone.0133862

Figure Lengend Snippet: A , Western blot analyses of Notch intercellular domain (NICD), hairy enhancer of split 1 (HES1) and endogenous control gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in cells isolated from human exocrine tissue. Whole cell lysates from CD133+ (+) and CD133-depleted (D) cells. Nuclear (N) and cytoplasmic (C) extracts from CD133+ cells. B , Volcano plot of Notch pathway gene mean ± SEM mRNA level (n = 3 exocrine cultures) differences in expression level from CD133+ cells compared to CD133D shown on X-axis as Log2 of fold difference. Significance determined by Student’s t-test shown on Y-axis as p value. Magenta vertical lines mark a 2-fold difference in expression. Blue horizontal line marks the significance cutoff (p<0.05). Selected gene names shown. Genes are: receptor tyrosine-protein kinase erbB-2 (ERBB2), frizzled class receptor 7 (FZD7), E1A binding protein p300 (EP300), MFNG O-fucosylpeptide 3-beta-N-acetylglucosaminyltransferase (MFNG), H19, imprinted maternally expressed transcript (H19), LIM domain only 2 (rhombotin-like 1) (LMO2), inhibitor of DNA binding 1 (ID1), hairy enhancer of split 4 (HES4), cyclin D1 (CCND1), matrix metallopeptidase 7 (MMP7), mastermind-like 2 (MAML2), jagged 1 (JAG1), Notch 2 (NOTCH2), hes-related family bHLH transcription factor with YRPW motif-like (HEYL), snail family zinc finger 2 (SNAI2), recombination signal binding protein for immunoglobulin kappa J region-like (RBPJL). C , Normalized mRNA expression level of neurogenin 3 (NGN3), HES1 and pancreas transcription factor 1 subunit alpha (PTF1A) in exocrine tissue after 4 days of culture in the presence of 20 μM Notch inhibitor DAPT. Results reported as mean ± SEM percent of levels in DMSO carrier control. mRNA levels normalized to the level of cyclophillin A. Significance determined by Student’s t-test, ***, p<0.001 (n = 3 exocrine cultures). D , Expression of NGN3 protein following treatment with 20 ∞M DAPT and 47 ∞M Notch agonist JAG-1 peptide (JAG-1). Mean ± SEM percent of DMSO carrier only control or 47 ∞M scrambled JAG-1 peptide, respectively indicated on Y-Axis. Significance determined by Student’s t-test, ***, p<0.001 (n = 3 exocrine cultures). E-H , Orthogonal analysis of colocalized HES1 and NGN3 in nuclei of exocrine tissue after 4 days of culture. Nuclei counterstained with Hoechst 33342 (H). E , Overlay of 3 channels. 0.5 ∞m confocal section. Scale bar is 50 ∞m. F-H , Higher magnification of crosshair region in all three channels shown at right. Scale bars are 20 ∞m. I , Coprecipitation of ID proteins with HES1. Whole cell lysate from exocrine tissue after 4 days of culture immunoprecipitated with antibody to HES1. ID1, 2 and 4 detected following SDS PAGE and western blotting. Predicted molecular weights of ID proteins (ID) and immunoglobulin heavy chain used for precipitation (HC) shown at right. Molecular weight marker positions shown at left in kDa.

Article Snippet: To investigate Notch signaling, tissue was treated with 20 μM N-[(3,5-Difluorophenyl)acetyl]-L-alanyl-2-phenyl]glycine-1,1-dimethylethyl ester (DAPT, TOCRIS Bioscience, Bristol, UK) resuspended in dimethylsulfoxide (DMSO) or with 47μM peptide corresponding to Jagged-1 amino acids 188–204 (JAG-1 peptide) or peptide with scrambled JAG-1 amino acids (Anaspec, Freemont, CA, USA) resuspended in water.

Techniques: Western Blot, Isolation, Expressing, Binding Assay, Immunoprecipitation, SDS Page, Molecular Weight, Marker