Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Effects of Sleep Deprivation (SD) on Rats via ERK1/2 Signaling Pathway

doi: 10.12659/MSM.913839

Figure Lengend Snippet: Effects of sleep deprivation (SD) on the expression of Orexin-A, OX1R, OX2R, PARP-1, ERK1/2, and p-ERK1/2 in rat hippocampal tissue. Total protein and total RNA were collected from hippocampal tissue using tissue extraction reagent. Western blot analysis was used to assess total protein, separation of protein, protein transference, incubation of primary antibody and secondary antibody, and coloration. We used qRT-PCR for cDNA synthesis and amplification. ( A ) Relative mRNA level was measured using qRT-PCR. ( B, C ) Relative protein level was evaluated using Western blot analysis. ANOVA with t tests was used to analyze data (* vs. control group; * p<0.05, ** p<0.01).

Article Snippet: TCSOX229, the OX2 receptor (OX2R) antagonist [ , ], and orexin-A were purchased from MCE (NJ, USA).

Techniques: Expressing, Extraction, Western Blot, Incubation, Quantitative RT-PCR, Amplification, Control

Journal: bioRxiv

Article Title: A Brain Reward Circuit Inhibited By Next-Generation Weight Loss Drugs

doi: 10.1101/2024.12.12.628169

Figure Lengend Snippet: a , Structure, naming convention, molecular formula, molecular weight (MW), half-life in humans, and species binding specificity of glucagon-like peptide-1 (GLP-1), liraglutide, danuglipron, and orforglipron. Amino acid sequences of liraglutide shown with its substitution and additions to GLP-1 highlighted in green and purple, respectively. b , Standard diet (SD) consumption over 2 hours post-administration of saline (Sal), liraglutide (Lira), vehicle (Veh), or danuglipron (Dan) ( n = 11 per injection, one-way ANOVA with Bonferroni correction, *** P <0.001). c , Schematic of the serine (TCA or S) to tryptophan (TGG or W) CRISPR-mediated substitution in Glp1r S33W (bottom) or in mouse Glp1r (top). d,e , Sanger sequencing chromatograms of ( d ) WT mouse Glp1r and ( e ) Glp1r S33W sequence, confirming the Glp1r S33W substitution in mice. f-h , Glucose tolerance test (GTT) on GLP1RAs. Comparison of blood glucose levels on ( f ) liraglutide (Lira), ( g ) danuglipron (Dan), ( h ) orforglipron (Orfo) followed by dextrose (Dex) in WT and Glp1r S33W mice ( n = 5-9 per injection, two-way ANOVA with Bonferroni correction, ** P <0.01; *** P <0.001). i-n , SD intake 1, 2, and 4 hours post-treatment of ( i, j ) Lira ( n = 9-11), ( k, l ) Dan ( n = 15-16), and ( m, n ) Orfo ( n = 8-9) and vehicle controls in WT and Glp1r S33W mice. o-t , High fat diet (HFD) intake 1, 2, and 4 hours post-treatment of ( o, p ) Lira ( n = 8-10), ( q, r ) Dan ( n = 14-16), and ( s, t ) Orfo ( n = 8-9) and their vehicle controls in WT and Glp1r S33W mice (two-way ANOVA with Bonferroni correction, * P <0.05; ** P <0.01; *** P <0.001). u , GTT measuring blood glucose levels following oral administration of danuglipron (oDan) and dextrose in Glp1r S33W and WT mice ( n = 6 per genotype, two-way ANOVA with Bonferroni correction, ** P <0.01; *** P <0.001). v-y , HFD intake 1, 2, and 4 hours after oral gavage of ( v ) danuglipron or vehicle and ( x ) orforglipron or saline ( n = 8-9, two-way ANOVA with Bonferroni correction, ** P <0.01; *** P <0.001). Comparison of the 4th hour between intraperitoneal (IP) and oral routes of administration with ( w ) danuglipron or ( y ) orforglipron (two-way ANOVA with Bonferroni correction, ** P <0.01; *** P <0.001). z , 7-day weight changes of mice on chronic HFD treated with orforglipron or saline daily at ZT6 ( n = 7 per injection, paired t-test, *** P <0.001). Data are represented as means ± SEM. * P <0.05; ** P <0.01; *** P <0.001. See Supplementary Table 1 for statistical details for all figures and Extended Data figures.

Article Snippet: Orforglipron powder (MedChemExpress, catalog no. HY-112185) was dissolved to 10mg/mL in Dimethyl sulfoxide (DMSO) and further diluted in 0.9% NaCl sterile saline to 0.1mg/mL.

Techniques: Molecular Weight, Binding Assay, Saline, Injection, CRISPR, Sequencing, Comparison

Journal: bioRxiv

Article Title: A Brain Reward Circuit Inhibited By Next-Generation Weight Loss Drugs

doi: 10.1101/2024.12.12.628169

Figure Lengend Snippet: GLP1RA activation across targeted GLP1R-expressing brain regions . a-d , GLP1R protein expression validated by a Glp1r-Cre;tdTomato mouse line and neuronal cFos activation 2 hours after danuglipron or liraglutide or 6 hours after orforglipron injection in WT and Glp1r S33W mice in the ( a ) DMH, ( b ) NTS, ( c ) AP, or ( d ) CeA. Scale bars = 200 µm. e-h , Quantification of cFos in the ( e ) DMH, ( f ) NTS, ( g ) AP and ( h ) CeA ( n = 3-4 per genotype, two-way ANOVA with Bonferroni correction, * P <0.05; ** P <0.01; *** P <0.001). i , Ratio of NTS/AP cFos activation in Glp1r S33W mice after danuglipron, orforglipron, and liraglutide ( n = 3-4 per injection, Kruskal-Wallis test, * P <0.05). j,k , Neuronal cFos activation in the NTS and AP 2 hours after danuglipron was administered to Glp1r S33W mice via ( j ) IP injection or ( k ) oral gavage. l , Quantification of cFos expression in the NTS (left) or AP (right) following danuglipron IP or oral delivery in Glp1r S33W mice ( n = 3 per delivery route, Welch’s t-test, * P <0.05). Data are represented as means ± SEM. * P <0.05; ** P <0.01; *** P <0.001.

Article Snippet: Orforglipron powder (MedChemExpress, catalog no. HY-112185) was dissolved to 10mg/mL in Dimethyl sulfoxide (DMSO) and further diluted in 0.9% NaCl sterile saline to 0.1mg/mL.

Techniques: Activation Assay, Expressing, Injection

Journal: bioRxiv

Article Title: A Brain Reward Circuit Inhibited By Next-Generation Weight Loss Drugs

doi: 10.1101/2024.12.12.628169

Figure Lengend Snippet: a , Representative image of the VTA from Glp1r-Cre mice injected with AAV- DIO- mGFP-2A-Synaptophysin-mRuby in the CeA; scale bar = 100 µm. CeA GLP1R fibers (red; pseudo-colored mGFP), synaptic terminals (green; pseudo-colored mRuby), and tyrosine hydroxylase-positive neurons (Th; blue, marking dopaminergic neurons) are shown, with a magnified view (right; scale bar = 50 µm). b , Schematic of proposed neural circuit from NTS → CeA → VTA → NAc. Arrows indicate neuron activation, blunted ends indicate neuron inhibition. Gray arrow from NTS → VTA indicates a known connection from NTS GLP-1 to VTA vGAT neurons . c , Schematic of genetically-encoded dopamine sensor, AAV-dLight1.3b, injection and fiber optic implant in the NAc of Glp1r S33W mice. d,g , Averaged Z-score traces showing dopamine release in the NAc in response to HFD following administration of ( d ) vehicle or danuglipron and ( g ) saline or orforglipron in Glp1r S33W mice. Traces are aligned to food retrieval time (t = 0) and averaged across five food trials per mouse. e-i , Quantified ( e,h ) area under the curve (AUC) for Z-scores and ( f,i ) maximum fluorescence Z-scores within the food retrieval window ( n = 9 for danuglipron, n = 7 for orforglipron, paired t-test, * P <0.05). j , Schematic of AAV-dLight1.3b injection and fiber optic implant into the NAc and AAV-DIO-hGLP1R injection into the CeA of Glp1r-Cre mice. k,n , Averaged Z-score traces showing dopamine release in the NAc in response to HFD following administration of ( k ) vehicle or danuglipron and ( n ) saline or orforglipron in CeA-hGLP1R mice. Traces are aligned to food retrieval time (t = 0) and averaged across five food trials per mouse. l- p , Quantification of ( l,o ) AUC for Z-scores and ( m,p ) maximum fluorescence Z-scores within the food retrieval window ( n = 8 for danuglipron, n = 7 for orforglipron, paired t-test, * P <0.05). Data are represented as means ± SEM. * P <0.05; ** P <0.01; *** P <0.001.

Article Snippet: Orforglipron powder (MedChemExpress, catalog no. HY-112185) was dissolved to 10mg/mL in Dimethyl sulfoxide (DMSO) and further diluted in 0.9% NaCl sterile saline to 0.1mg/mL.

Techniques: Injection, Activation Assay, Inhibition, Saline, Fluorescence

Journal: bioRxiv

Article Title: Hypoxia-induced Complement Component 3 Promotes Aggressive Tumor Growth in the Glioblastoma Microenvironment

doi: 10.1101/2024.01.28.577617

Figure Lengend Snippet: A-B. Immunofluorescent detection of C3 (red), Nestin (yellow), Nuclei (DAPI, blue), GFAP (green) or Olig2 (cyane) in the perivascular (A) or hypoxic (B) niches of murine RCAS-PDGFB- and RCAS-shp53-induced gliomas. C-D. Hallmark_Hypoxia (C) and Hallmark_Complement (D) signatures mapped on spatially resolved transcriptomics from human GBM samples, displayed with Z-score. E. Spatial correlation between Hallmark_Hypoxia and Hallmark_Complement in one representative tumor (UKF242). P values were corrected for multiple hypothesis testing using the Holm method. F. Distribution of R-values for the correlation between Hallmark_Hypoxia and Hallmark_Complement in a total of 19 human GBM tissue samples with an average correlation score of R=0.54 G-H. Pearson correlation coefficient between Hallmark_Complement and Hallmark_Hypoxia signatures in the TCGA GBMLGG (G) or TCGA GBM (H) dataset. I-K. Expression of CA9, C3, and C3AR1 mRNA in human primary astrocytes (n=3), HMC3 microglia (n=4), or primary human glioma cells U3082MG (n=3), U3065MG (n=3) or U3084MG (n=3) in response to normoxia (21% O2), hypoxia (1% O2) or severe hypoxia (0.1% O2). Error bars represent SEM. *, P < 0.05, **, P < 0.01, or ***, P< 0,001. Statistical tests were one-way ANOVA, with Tukey post-hoc test, or unpaired t-test in case of two sample comparisons.

Article Snippet: The spheres were grown until visible spheres were formed (up to 14 days) with control or treatments with 50 and 250nM C3aR antagonist (Santa Cruz Biotechnology, SB290157).

Techniques: Expressing

Journal: bioRxiv

Article Title: Hypoxia-induced Complement Component 3 Promotes Aggressive Tumor Growth in the Glioblastoma Microenvironment

doi: 10.1101/2024.01.28.577617

Figure Lengend Snippet: A. C3AR1 expression of Pan-Cancer TCGA data of common cancer types (n=33). B. C3AR1 expression in relation to glioma grade as analyzed in TCGA data. C. C3AR1 expression in IDH wildtype glioma compared to IDH mutant with or without 1p/19q codeletion (Tukey post-hoc test) as analyzed in TCGA data. D. C3AR1 expression in GBM compared to non-tumor as analyzed in TCGA data. E. Kaplan-Meier curve showing survival of glioma patients with either high (red) or low (blue) C3AR1 expression based on TCGA data. F . Kaplan-Meier curve showing survival of IDH-wildtype GBM with high (red) or low (blue) C3AR1 expression based on TCGA data. G. UMAP displaying C3AR1 expression in single cell sequencing data from 26 independent datasets compiled in GBmap . H. C3AR1 expression of malignant cells divided into C3AR1-expressing or non-expressing cells. I. Geneset enrichment analysis of the C3AR1-expressing malignant cells. Red colored bars indicate significant Benjamini-Hochberg adjusted P values (padj < 0.05). J. Log-fraction plot of a combination of independent extreme limiting dilution sphere formation assays (n=4) of U3082MG glioma cells treated with SB290157. *, P < 0.05, **, P < 0.01, or ***, P< 0,001. One-way ANOVA, or unpaired T-test (in case of comparison between two groups) with Tukey post-hoc test.

Article Snippet: The spheres were grown until visible spheres were formed (up to 14 days) with control or treatments with 50 and 250nM C3aR antagonist (Santa Cruz Biotechnology, SB290157).

Techniques: Expressing, Mutagenesis, Sequencing, Comparison

Journal: Neoplasia (New York, N.Y.)

Article Title: Targeting PAR1 activation in JAK2V617F-driven philadelphia-negative myeloproliferative neoplasms: Unraveling its role in thrombosis and disease progression

doi: 10.1016/j.neo.2025.101153

Figure Lengend Snippet: Relative PAR1 gene expression analysis of (A) mononuclear cells ( p = 0.0005, Mann-Whitney U), (B) erythro‑myeloid restricted progenitors (EMPs, CD34+CD133-) ( p = 0.0008, Mann-Whitney U), (C) multipotent progenitor cells (MPPs, CD133+CD34+) ( p = 0.706, Mann-Whitney U), (D) hematopoietic and endothelial progenitor cells (HSC/ EPC, CD133-CD34-) ( p = 0.0002, Mann-Whitney U), (E) endothelial progenitor cells (EPCs, CD34-CD133+) ( p = 0.0064, Mann-Whitney U) in samples from 13 patients with myeloproliferative neoplasms and 5 control samples of cord blood. In each figure, the standard deviation of the mean (SD) is shown with a bar.

Article Snippet: The MNCs and/or CD45 - CD34 +/depleted cells of MPN patients or healthy volunteers were incubated with the combination of varying treatments including thrombin (Sigma Aldrich, St. Louis, Missouri, USA), selective PAR1 antagonist, Vorapaxar (SCH530348, MedChemExpress, Monmouth Junction, NJ, USA) and JAK2 inhibitor, Ruxolitinib (sc-364729, Santa Cruz Biotechnology, Dallas, Texas, USA) for the analysis.

Techniques: Gene Expression, MANN-WHITNEY, Control, Standard Deviation

Journal: Neoplasia (New York, N.Y.)

Article Title: Targeting PAR1 activation in JAK2V617F-driven philadelphia-negative myeloproliferative neoplasms: Unraveling its role in thrombosis and disease progression

doi: 10.1016/j.neo.2025.101153

Figure Lengend Snippet: JAK2V617F allele status of Mononuclear Cells and PAR1 sorted populations of Ph- MPN samples.

Article Snippet: The MNCs and/or CD45 - CD34 +/depleted cells of MPN patients or healthy volunteers were incubated with the combination of varying treatments including thrombin (Sigma Aldrich, St. Louis, Missouri, USA), selective PAR1 antagonist, Vorapaxar (SCH530348, MedChemExpress, Monmouth Junction, NJ, USA) and JAK2 inhibitor, Ruxolitinib (sc-364729, Santa Cruz Biotechnology, Dallas, Texas, USA) for the analysis.

Techniques:

Journal: Neoplasia (New York, N.Y.)

Article Title: Targeting PAR1 activation in JAK2V617F-driven philadelphia-negative myeloproliferative neoplasms: Unraveling its role in thrombosis and disease progression

doi: 10.1016/j.neo.2025.101153

Figure Lengend Snippet: PAR1 cell surface expression analysis of MPN cells (4 MPN patients with monoallelic JAK2V617F) (A) fluorescence-activated cell sorting plot with CD34 and PAR1 expression, (B and C) cell sorting according to CD34 and PAR1 expression that lack of CD45 expression. (D) EPCR cell surface expression analysis of MPN cells with CD34 expression.

Article Snippet: The MNCs and/or CD45 - CD34 +/depleted cells of MPN patients or healthy volunteers were incubated with the combination of varying treatments including thrombin (Sigma Aldrich, St. Louis, Missouri, USA), selective PAR1 antagonist, Vorapaxar (SCH530348, MedChemExpress, Monmouth Junction, NJ, USA) and JAK2 inhibitor, Ruxolitinib (sc-364729, Santa Cruz Biotechnology, Dallas, Texas, USA) for the analysis.

Techniques: Expressing, Fluorescence, FACS

Journal: Neoplasia (New York, N.Y.)

Article Title: Targeting PAR1 activation in JAK2V617F-driven philadelphia-negative myeloproliferative neoplasms: Unraveling its role in thrombosis and disease progression

doi: 10.1016/j.neo.2025.101153

Figure Lengend Snippet: The effect of thrombin on PAR1 expression was evaluated in (A) CD34 +/depleted cells who had lack of CD45 expression (B) the inhibitory effects of vorapaxar were tested in PAR1 cell surface expression (C) and on the phosphorylation of STAT-5B. GraphPad Prism 8.0 software was used for the statistical analysis. (Friedman Test; mean ± SEM, * P < 0.05, ** P < 0.01, ** P < 0.001).

Article Snippet: The MNCs and/or CD45 - CD34 +/depleted cells of MPN patients or healthy volunteers were incubated with the combination of varying treatments including thrombin (Sigma Aldrich, St. Louis, Missouri, USA), selective PAR1 antagonist, Vorapaxar (SCH530348, MedChemExpress, Monmouth Junction, NJ, USA) and JAK2 inhibitor, Ruxolitinib (sc-364729, Santa Cruz Biotechnology, Dallas, Texas, USA) for the analysis.

Techniques: Expressing, Software

Journal: Neoplasia (New York, N.Y.)

Article Title: Targeting PAR1 activation in JAK2V617F-driven philadelphia-negative myeloproliferative neoplasms: Unraveling its role in thrombosis and disease progression

doi: 10.1016/j.neo.2025.101153

Figure Lengend Snippet: The effect of PAR1 activation with thrombin in JAK2 pathway. Mononuclear cells obtained from MPN patients (n: 11) and healthy controls (n: 11) were cultured overnight in RPMI 1640 serum-free conditions and then treated with Ruxolitinib (300 nm, 3 h (monoallelic JAK2V617F) or 1.5 h (biallelic JAK2V617F and wild type), thrombin (1U, 1 h), vorapaxar (80uM, 1 h) to perform relative PAR1 gene expression analysis (A-I). In patients with monoallelic JAK2V617F (n: 4), biallelic JAK2V617F (n: 5) and wild type (n: 2), the effects of thrombin, vorapaxar and ruxolitinib administration on relative PAR1 gene expression were analyzed (J).

Article Snippet: The MNCs and/or CD45 - CD34 +/depleted cells of MPN patients or healthy volunteers were incubated with the combination of varying treatments including thrombin (Sigma Aldrich, St. Louis, Missouri, USA), selective PAR1 antagonist, Vorapaxar (SCH530348, MedChemExpress, Monmouth Junction, NJ, USA) and JAK2 inhibitor, Ruxolitinib (sc-364729, Santa Cruz Biotechnology, Dallas, Texas, USA) for the analysis.

Techniques: Activation Assay, Cell Culture, Gene Expression

Journal: Neoplasia (New York, N.Y.)

Article Title: Targeting PAR1 activation in JAK2V617F-driven philadelphia-negative myeloproliferative neoplasms: Unraveling its role in thrombosis and disease progression

doi: 10.1016/j.neo.2025.101153

Figure Lengend Snippet: The cell surface PAR1 expression pattern evaluated after treated MNC cells with thrombin (1U), vorapaxar (80uM), ruxolitinib (300 nm) healthy controls (n:11, p < 0.0001) (A), MPN patients (n:11, p < 0.0001) (B) and MPN patients have history of thrombosis (n:4, p:0.001) (C). Results are performed on two separate occasions. GraphPad Prism 8.0 software was used for the statistical analysis of cell surface protein expression data. (Friedmann Test).

Article Snippet: The MNCs and/or CD45 - CD34 +/depleted cells of MPN patients or healthy volunteers were incubated with the combination of varying treatments including thrombin (Sigma Aldrich, St. Louis, Missouri, USA), selective PAR1 antagonist, Vorapaxar (SCH530348, MedChemExpress, Monmouth Junction, NJ, USA) and JAK2 inhibitor, Ruxolitinib (sc-364729, Santa Cruz Biotechnology, Dallas, Texas, USA) for the analysis.

Techniques: Expressing, Software

Journal: Neoplasia (New York, N.Y.)

Article Title: Targeting PAR1 activation in JAK2V617F-driven philadelphia-negative myeloproliferative neoplasms: Unraveling its role in thrombosis and disease progression

doi: 10.1016/j.neo.2025.101153

Figure Lengend Snippet: The responses of MPN cells to PAR1 inhibition by detection of changes in PAR pathway. (A) Jak2V617F-induced PAR regulated genes and (C) trombin-PAR-coagulation suppressed genes affected >2-fold in the PAR pathway in mononuclear cells of MPN patients (2 patients with monoallelic JAK2V617F (p26 and p:29), 2 patients with biallelic JAK2V617F (p:64 and p:68) and no mutation carrying one patient (p: 35)) by PAR1 inhibition. (B) PAR pathway-related genes downregulated >2-fold in one MPN patient with history of thrombosis (p:26).

Article Snippet: The MNCs and/or CD45 - CD34 +/depleted cells of MPN patients or healthy volunteers were incubated with the combination of varying treatments including thrombin (Sigma Aldrich, St. Louis, Missouri, USA), selective PAR1 antagonist, Vorapaxar (SCH530348, MedChemExpress, Monmouth Junction, NJ, USA) and JAK2 inhibitor, Ruxolitinib (sc-364729, Santa Cruz Biotechnology, Dallas, Texas, USA) for the analysis.

Techniques: Inhibition, Coagulation, Mutagenesis

Journal: International Journal of Molecular Sciences

Article Title: Structural Characteristics of Polysaccharide GP2a in Gardenia jasminoides and Its Immunomodulatory Effect on Macrophages

doi: 10.3390/ijms231911279

Figure Lengend Snippet: Fourier-transform infrared (FTIR) spectrum of GP2a. Bands positions are indicated in cm −1 .

Article Snippet: Thereafter, a culture medium with GP2a at various concentrations (7.5–300 µg/mL) was added and incubated for 24, 48, or 72 h. Meanwhile, untreated cells were used as blank control and the cells treated with 2 µg/mL LPS (MedChemExpress) were used as positive control.

Techniques: Fourier Transform Infrared Spectroscopy

Journal: International Journal of Molecular Sciences

Article Title: Structural Characteristics of Polysaccharide GP2a in Gardenia jasminoides and Its Immunomodulatory Effect on Macrophages

doi: 10.3390/ijms231911279

Figure Lengend Snippet: Plots of molecular weight distribution ( A ) and molecular conformation ( B ) of GP2a. ( A ) Superimposed chromatograms of GP2a obtained by size exclusion chromatography connected with multi-angle laser light scattering and refractive index (RI) detectors; the RI trace shows the signals collected by the RI detector; the molar mass trace was fitted by laser light scattering signals and RI signals, indicating molecular weight distribution. ( B ) Log–log plot of r g versus Mw; RMS—root mean square.

Article Snippet: Thereafter, a culture medium with GP2a at various concentrations (7.5–300 µg/mL) was added and incubated for 24, 48, or 72 h. Meanwhile, untreated cells were used as blank control and the cells treated with 2 µg/mL LPS (MedChemExpress) were used as positive control.

Techniques: Molecular Weight, Size-exclusion Chromatography, Refractive Index

Journal: International Journal of Molecular Sciences

Article Title: Structural Characteristics of Polysaccharide GP2a in Gardenia jasminoides and Its Immunomodulatory Effect on Macrophages

doi: 10.3390/ijms231911279

Figure Lengend Snippet: Ion chromatogram of standard monosaccharides ( A ) and GP2a hydrolysate ( B ). Peaks in ( A ): 1 is fucose (Fuc), 2 is arabinose (Ara), 3 is rhamnose (Rha), 4 is galactose (Gal), 5 is glucose (Glc), 6 is xylose (Xyl), 7 is mannose (Man), 8 is fructose (Fru), 9 is ribose (Rib), 10 is galacturonic acid (GalA), 11 is guluronic acid (GulA), 12 is glucuronic acid (GlcA), and 13 is mannuronic acid (ManA). Peaks in ( B ): 1 is Fuc, 2 is Ara, 3 is Rha, 4 is Gal, 5 is Glc, 6 is Xyl, 7 is Man, 8 is GalA, and 9 is GlcA.

Article Snippet: Thereafter, a culture medium with GP2a at various concentrations (7.5–300 µg/mL) was added and incubated for 24, 48, or 72 h. Meanwhile, untreated cells were used as blank control and the cells treated with 2 µg/mL LPS (MedChemExpress) were used as positive control.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Structural Characteristics of Polysaccharide GP2a in Gardenia jasminoides and Its Immunomodulatory Effect on Macrophages

doi: 10.3390/ijms231911279

Figure Lengend Snippet: Monosaccharide composition of GP2a.

Article Snippet: Thereafter, a culture medium with GP2a at various concentrations (7.5–300 µg/mL) was added and incubated for 24, 48, or 72 h. Meanwhile, untreated cells were used as blank control and the cells treated with 2 µg/mL LPS (MedChemExpress) were used as positive control.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Structural Characteristics of Polysaccharide GP2a in Gardenia jasminoides and Its Immunomodulatory Effect on Macrophages

doi: 10.3390/ijms231911279

Figure Lengend Snippet: Total ion chromatograms (TIC) of GP2a derivatives. ( A ) H/D, singly deuterated sample, referring to the carboxyl groups prereduced using sodium borohydride (NaBH 4 ) and secondary reduced using sodium borodeuteride (NaBD 4 ) after methylation. ( B ) D/D, doubly deuterated sample, referring to the carboxyl groups reduced using NaBD 4 before and after methylation. +EI: electron bombardment ion source.

Article Snippet: Thereafter, a culture medium with GP2a at various concentrations (7.5–300 µg/mL) was added and incubated for 24, 48, or 72 h. Meanwhile, untreated cells were used as blank control and the cells treated with 2 µg/mL LPS (MedChemExpress) were used as positive control.

Techniques: Methylation

Journal: International Journal of Molecular Sciences

Article Title: Structural Characteristics of Polysaccharide GP2a in Gardenia jasminoides and Its Immunomodulatory Effect on Macrophages

doi: 10.3390/ijms231911279

Figure Lengend Snippet: Methylation analysis result of GP2a.

Article Snippet: Thereafter, a culture medium with GP2a at various concentrations (7.5–300 µg/mL) was added and incubated for 24, 48, or 72 h. Meanwhile, untreated cells were used as blank control and the cells treated with 2 µg/mL LPS (MedChemExpress) were used as positive control.

Techniques: Methylation, Molecular Weight

Journal: International Journal of Molecular Sciences

Article Title: Structural Characteristics of Polysaccharide GP2a in Gardenia jasminoides and Its Immunomodulatory Effect on Macrophages

doi: 10.3390/ijms231911279

Figure Lengend Snippet: Scanning electron microscopy (SEM) images of GP2a at ( A ) 100×; ( B ) 250×; ( C ) 500×; ( D ) 1000×.

Article Snippet: Thereafter, a culture medium with GP2a at various concentrations (7.5–300 µg/mL) was added and incubated for 24, 48, or 72 h. Meanwhile, untreated cells were used as blank control and the cells treated with 2 µg/mL LPS (MedChemExpress) were used as positive control.

Techniques: Electron Microscopy

Journal: International Journal of Molecular Sciences

Article Title: Structural Characteristics of Polysaccharide GP2a in Gardenia jasminoides and Its Immunomodulatory Effect on Macrophages

doi: 10.3390/ijms231911279

Figure Lengend Snippet: Nuclear magnetic resonance (NMR) spectra of GP2a. ( A ) 1 H-NMR spectrum; ( B ) 13 C-NMR spectrum; ( C ) Heteronuclear single quantum relation (HSQC) spectrum; ( D ) Correlated spectroscopy (COSY) spectrum; ( E ) Heteronuclear multiple bond correlation (HMBC) spectrum; ( F ) Nuclear Overhauser effect spectroscopy (NOESY) spectrum.

Article Snippet: Thereafter, a culture medium with GP2a at various concentrations (7.5–300 µg/mL) was added and incubated for 24, 48, or 72 h. Meanwhile, untreated cells were used as blank control and the cells treated with 2 µg/mL LPS (MedChemExpress) were used as positive control.

Techniques: Nuclear Magnetic Resonance, Spectroscopy

Journal: International Journal of Molecular Sciences

Article Title: Structural Characteristics of Polysaccharide GP2a in Gardenia jasminoides and Its Immunomodulatory Effect on Macrophages

doi: 10.3390/ijms231911279

Figure Lengend Snippet: 1 H NMR and 13 C NMR chemical shifts of GP2a recorded in D 2 O.

Article Snippet: Thereafter, a culture medium with GP2a at various concentrations (7.5–300 µg/mL) was added and incubated for 24, 48, or 72 h. Meanwhile, untreated cells were used as blank control and the cells treated with 2 µg/mL LPS (MedChemExpress) were used as positive control.

Techniques: Residue

Journal: International Journal of Molecular Sciences

Article Title: Structural Characteristics of Polysaccharide GP2a in Gardenia jasminoides and Its Immunomodulatory Effect on Macrophages

doi: 10.3390/ijms231911279

Figure Lengend Snippet: Possible repeat unit structure of GP2a.

Article Snippet: Thereafter, a culture medium with GP2a at various concentrations (7.5–300 µg/mL) was added and incubated for 24, 48, or 72 h. Meanwhile, untreated cells were used as blank control and the cells treated with 2 µg/mL LPS (MedChemExpress) were used as positive control.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Structural Characteristics of Polysaccharide GP2a in Gardenia jasminoides and Its Immunomodulatory Effect on Macrophages

doi: 10.3390/ijms231911279

Figure Lengend Snippet: Cell viability of RAW 264.7 cells.

Article Snippet: Thereafter, a culture medium with GP2a at various concentrations (7.5–300 µg/mL) was added and incubated for 24, 48, or 72 h. Meanwhile, untreated cells were used as blank control and the cells treated with 2 µg/mL LPS (MedChemExpress) were used as positive control.

Techniques: Control, Positive Control

Journal: International Journal of Molecular Sciences

Article Title: Structural Characteristics of Polysaccharide GP2a in Gardenia jasminoides and Its Immunomodulatory Effect on Macrophages

doi: 10.3390/ijms231911279

Figure Lengend Snippet: Effects of GP2a on nitric oxide (NO) production in RAW 264.7 cells. Untreated cells were used as blank control; Cells treated with different GP2a concentrations (120, 240, and 300 µg/mL) were used as samples; Cells treated with lipopolysaccharide (LPS) (2 µg/mL) were used as positive control. Letter a represents significant difference compared with the blank control ( p < 0.05, p < 0.01); letter b represents significant difference compared with the positive control ( p < 0.05, p < 0.01).

Article Snippet: Thereafter, a culture medium with GP2a at various concentrations (7.5–300 µg/mL) was added and incubated for 24, 48, or 72 h. Meanwhile, untreated cells were used as blank control and the cells treated with 2 µg/mL LPS (MedChemExpress) were used as positive control.

Techniques: Control, Positive Control

Journal: International Journal of Molecular Sciences

Article Title: Structural Characteristics of Polysaccharide GP2a in Gardenia jasminoides and Its Immunomodulatory Effect on Macrophages

doi: 10.3390/ijms231911279

Figure Lengend Snippet: Effects of GP2a on cytokine production in RAW 264.7 cells. ELISA detection of tumor necrosis factor-α (TNF-α) ( A ), interferon (IFN)-γ ( B ), interleukin (IL)-1β ( C ), IL-6 ( D ), and granulocyte macrophage colony stimulating factor (GM-CSF) ( E ). Untreated cells were used as blank control; cells treated with different GP2a concentrations (120, 240, and 300 µg/mL) were used as sample; cells treated with lipopolysaccharide (LPS) (2 µg/mL) were used as positive control. Letter a represents significant difference compared with the blank control ( p < 0.05, p < 0.01); letter b represents significant difference compared with the positive control ( p < 0.05, p < 0.01).

Article Snippet: Thereafter, a culture medium with GP2a at various concentrations (7.5–300 µg/mL) was added and incubated for 24, 48, or 72 h. Meanwhile, untreated cells were used as blank control and the cells treated with 2 µg/mL LPS (MedChemExpress) were used as positive control.

Techniques: Enzyme-linked Immunosorbent Assay, Control, Positive Control

Journal: Nature Communications

Article Title: GLP-1R associates with VAPB and SPHKAP at ERMCSs to regulate β-cell mitochondrial remodelling and function

doi: 10.1038/s41467-025-66115-x

Figure Lengend Snippet: a EGFP-VAPB co-IP with SNAP/FLAG-hGLP-1R in vehicle (Veh) versus exendin-4 (Ex-4)-stimulated INS-1 832/3 SNAP/FLAG-hGLP-1R cells co-expressing control pcDNA3.1+ versus dominant negative dynamin (Dyn)1/2 K44A; n = 1. b Veh-subtracted Ex-4- versus exendin-F1 (Ex-F1)-induced BRET kinetic traces (left) and corresponding AUCs (right) of SNAP/FLAG-hGLP-1R-NLuc – Venus-VAPB interactions in INS-1 832/3 cells; n = 3 biologically independent experiments, p value as indicated by two-tailed paired t -test. c (top), Confocal microscopy analysis of SNAP/FLAG-hGLP-1R localisation in mouse primary islets transduced with SNAP/FLAG-hGLP-1R adenoviruses under Veh conditions or following 5-minute stimulation with the indicated agonists: Ex-4, Ex-F1, exendin-D3 (Ex-D3), tirzepatide (Tirz) and semaglutide (Sema) used at 100 nM; orforglipron (Orf) and danuglipron (Dan) used at 5 μM; size bars, 20 μm; c (bottom), Quantification of residual surface SNAP/FLAG-hGLP-1R levels in islets from above; n = 3 biologically independent experiments. d Quantification of VAPB co-IP with SNAP/FLAG-hGLP-1R in INS-1 832/3 SNAP/FLAG-hGLP-1R cells stimulated for 5 minutes with the indicated agonists; n = 3 biologically independent experiments. e VAPB-localised cAMP generation (measured by FluoSTEP) in response to 100 nM Ex-4 versus Ex-F1 stimulation in INS-1 832/3 cells expressing pcDNA3-GFP11(x7)-VAPB + pcDNA3.1(+)-FluoSTEP-ICUE; maximal responses [to forskolin (FSK) + 3-Isobutyl-1-methylxanthine (IBMX)] shown; kinetic traces (left) and corresponding AUCs for the agonist-stimulated period (right) shown; n = 5 biologically independent experiments, p value as indicated by two-tailed paired t -test. f (top), Ex-4 versus Ex-F1-induced global PKA activity, measured with the global PKA biosensor pcDNA3.1(+)- ExRai-AKAR2 in INS-1 832/3 cells; kinetic traces (left) and corresponding AUCs (right) shown; n = 3 biologically independent experiments; p value as indicated by two-tailed paired t -test. f (bottom), Ex-4 versus Ex-F1-induced mitochondrial PKA activity, measured with the mitochondrial PKA biosensor pcDNA3-mitoExRai-AKAR2 in INS-1 832/3 cells; kinetic traces (left) and corresponding AUCs (right) shown; n = 3 biologically independent experiments, p value as indicated by two-tailed paired t -test. g , Mitochondrial over global PKA activity, calculated as AUC ratio from data in ( f ); n = 3 biologically independent experiments, p value as indicated by two-tailed paired t -test. Data are mean ± SEM.

Article Snippet: Custom analogues (exendin-F1 and exendin-D3) were from WuXi AppTec, China, and danuglipron and orforglipron were purchased from MedChemExpress.

Techniques: Co-Immunoprecipitation Assay, Expressing, Control, Dominant Negative Mutation, Two Tailed Test, Confocal Microscopy, Transduction, Activity Assay