annexin a1 protein  (MedChemExpress)

Journal: Signal Transduction and Targeted Therapy

Article Title: Single-cell transcriptional dissection illuminates an evolution of immunosuppressive microenvironment during pancreatic ductal adenocarcinoma metastasis

doi: 10.1038/s41392-025-02265-0

Figure Lengend Snippet: Evolution of PDAC metastasis accompanies the function loss of DC. a UMAP plot represents the sub-clustered DCs from all biopsies, with cells color-coded according to cell type for clear differentiation. b A heatmap shows the DEGs for each DC subtype, with representative marker genes highlighted and color indicating the normalized z-score. c A box plot illustrates the expression of marker genes across different cell types, including pDC ( LILRA4 + , GZMB + ), mDC ( LAMP3 + ), cDC1 ( CLEC9A + ), cDC2 ( CD1C + , CLEC10A + ), and immature DC ( CD1A + ), with color denoting gene group affiliation. d A bar plot displays the proportion of each DC subtype within the immune cell population. e A heatmap represents the differentially enriched gene sets for each DC subtype. f The dot plot illustrates the expression levels of ligand-receptor pairs, specifically ANXA1_FPR1/2, between TECs/NECs and DC subtypes across tissues. Color intensity reflects the mean expression of the ligand and receptor genes, while dot size corresponds to the statistical significance of the interactive molecular pairs. g Violin plots present the expression levels of FPR1 on cDCs across different tissues (left), FPR3 on cDCs (middle), and ANXA1 on TECs/NECs (right). h Representative images of mIHC staining with HD tissue, primary_NM tumor, primary_HM tumor, and HM lesion. DAPI (blue), CD11c (green), CK (cyan), cleaved-caspase3 (red), TNF-α (purple), ANXA1 (yellow). Scale bars, 200 μm, or 100 μm

Article Snippet: The cells were then treated with indicated doses (0, 5, 10, and 20 μg/ml) of ANXA1 protein (MedChemExpress, HY- P72078 ) for 24 h, based on previous studies demonstrating the effective modulation of DC function within this concentration range., , After incubation, cell apoptosis was detected with Annexin V-FITC/PI Apoptosis Detection Kit according to the manufacturer’s instructions (Beijing 4A Biotech Co., Ltd., #FXP018).

Techniques: Marker, Expressing, Staining

Journal: Signal Transduction and Targeted Therapy

Article Title: Single-cell transcriptional dissection illuminates an evolution of immunosuppressive microenvironment during pancreatic ductal adenocarcinoma metastasis

doi: 10.1038/s41392-025-02265-0

Figure Lengend Snippet: Validation of ANXA1 function in vitro and in vivo. The apoptosis of BMDC cells treated with indicated doses (0, 5, 10, 20 μg/ml) of ANXA1 was evaluated by Annexin V-FITC/PI based flow cytometry ( a ) and the results were quantified ( b ). Proteins indicative of apoptosis pathway activity, including MCL-1, BCL-2, and cleaved caspase3, were detected in BMDC ( c ) and DC2.4 ( d ) cells treated with 10 μg/ml ANXA1 for 24 h. e , f The PDAC xenograft model was established by subcutaneous injection of KPC cells with or without manipulation of the expression of ANXA1. Anti-PD1 treatment started when tumor volume achieved ≈80 mm 3 . The average tumor growth curves were profiled during treatment ( d ), and the tumor weight was recorded at the end of the treatment ( e ). g , h Mouse PDAC orthotopic models were established by injecting luciferase-tagged KPC cells (KPC-Luc) with ANXA1 overexpression, ANXA1 knockdown, or empty vector into the pancreas tissues of mice. Anti-PD-1 treatment (2.5 mg/kg, i.v.) was administered every 6 days after 5 days of the tumor inoculation. The tumor growth was visualized ( g ) and quantified ( h ) by bioluminescence imaging system 20 days after tumor inoculation. i Whole slide immunofluorescence staining of CD8 and CD11c to assess the infiltration of T cells and DCs in subcutaneous PDAC tumors. Scale bar 200 μm or 50 μm. j , k Quantification of the densities of CD8 + T cells ( h ) and CD11c + DCs ( i ) in each group from ( i )

Article Snippet: The cells were then treated with indicated doses (0, 5, 10, and 20 μg/ml) of ANXA1 protein (MedChemExpress, HY- P72078 ) for 24 h, based on previous studies demonstrating the effective modulation of DC function within this concentration range., , After incubation, cell apoptosis was detected with Annexin V-FITC/PI Apoptosis Detection Kit according to the manufacturer’s instructions (Beijing 4A Biotech Co., Ltd., #FXP018).

Techniques: Biomarker Discovery, In Vitro, In Vivo, Flow Cytometry, Activity Assay, Injection, Expressing, Luciferase, Over Expression, Knockdown, Plasmid Preparation, Imaging, Immunofluorescence, Staining

Journal: Endocrine

Article Title: Mesenchymal stromal cells and their secretory products reduce the inflammatory crosstalk between islets and endothelial cells.

doi: 10.1007/s12020-024-03975-1

Figure Lengend Snippet: Fig. 3 ANXA1 reduces TNF- induced primary HUVEC activation. Confocal micrographs showing representative images of Z-stack projected HUVEC monolayer images comprised of 7–15 × 0.68 µm slices, 20× magnification. HUVEC monolayers were exposed to 100 U/ml TNF-α alone (a), or to 100 U/ml TNF in the presence of 5 nM ANXA1 (b), for 24 h. Green and red indicate ICAM-1 and VCAM-1 immunostaining, respectively. The percentage of HUVECs immunopositive for VCAM-1 alone (c), ICAM-1 alone (d), or dual positive for both V-CAM1 and I-CAM1 (e) was assessed using HALO® v3.3, Cytonuclear FL V2.0.12 module (Indica Labs) image analysis software. 14-15 fields of view per condition assessed, *p < 0.05, **p < 0.01 vs. HUVECs exposed to 100 U/ml TNF without ANXA1 treatment. The p values were calculated using Student’s t-test

Article Snippet: After washing with RPMI-1640 medium, islets were picked into groups of 80-100 for culture alone, with MSCs, with recombinant ANXA1 alone (R & D Systems, Abingdon, UK), or with a defined cocktail of MSC-secretory factors (recombinant ANXA1 (5nmol/L), recombinant mouse SDF-1/CXCL12 (10 nmol/L), and recombinant mouse C3a (10nmol/L) [24].

Techniques: Activation Assay, Immunostaining, Software

Journal: Small (Weinheim an der Bergstrasse, Germany)

Article Title: Laminin-Augmented Decellularized Extracellular Matrix Ameliorating Neural Differentiation and Neuroinflammation in Human Mini-Brains.

doi: 10.1002/smll.202308815

Figure Lengend Snippet: Figure 4. ANXA1 in P-BdECM promotes microglial inflammation. A) Overview of microglial immune activity characterization. B) Representative fluores- cence images of ROS, CD86, iNOS, and NO of SV40 human microglia grown in Matrigel (MTG) and hybrid hydrogel under normal conditions (-LPS). C–F) Quantification of (C) ROS (n = 25), (D) CD86 (n = 30), (E) iNOS (n = 30), and (F) NO (n = 44). G) Representative fluorescence images showing colocalization of microglia with fluorescein isothiocyanate (FITC)-labeled zymosan particles (left), FITC-labeled monomeric amyloid beta (mA𝛽) (mid- dle), and TREM2-staining (+mA𝛽) (right); cell outlines are shown by white dashed lines. H–J) Quantification of (H) zymosan phagocytosis in microglia (n = 50), (I) colocalization of microglia with mA𝛽(n = 40), (J) TREM2 staining (+mA𝛽) (n = 30). All data are presented as means ± SEM. Statistical analyses were performed using one-way ANOVA with Turkey’s multiple comparisons test (C,D,E,F,H,I,J); *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Scale bars, 100 μm (B; G, TREM2); 10 μm (G, zymosan); 20 μm (G, mA𝛽). All experiments were performed in biological triplicates.

Article Snippet: For the ANXA1 augmentation, 2 mg mL−1 of Matrigel was supplemented with human recombinant ANXA1 (hrANXA1) (3770-AN-050, R&D Systems) to reach the final concentration of 0.74 μg mL−1 hrANXA1.

Techniques: Activity Assay, Labeling, Staining

Journal: Small (Weinheim an der Bergstrasse, Germany)

Article Title: Laminin-Augmented Decellularized Extracellular Matrix Ameliorating Neural Differentiation and Neuroinflammation in Human Mini-Brains.

doi: 10.1002/smll.202308815

Figure Lengend Snippet: Figure 5. ANAX1 in P-BdECM promotes astrocytic inflammation. A) Overview of the astrocytic immune activity characterization. B) Representative fluorescence images of GFAP staining of astrocytes grown in Matrigel (MTG), P-BdECM (BdECM), hybrid, and ANXA1-augmented MTG (MTG+ANXA1); WRW4, a FPR2 antagonist. C) Quantitative analysis for GFAP intensity (n ≥25). D) Representative fluorescence images for intracellular ROS in astrocytes. E) Quantitative analysis for ROS intensity. All data are presented as means ± SEM. Statistical analyses were performed using one-way ANOVA with Dunnett’s multiple comparisons test (n ≥25). (C,E); *p < 0.05, ****p < 0.0001, ####p < 0.0001. In (C,E), “*” indicates comparisons with the (MTG, -LPS) group, “#” indicates comparisons with the (MTG, +LPS) group. Scale bars, 100 μm (B,D). All experiments were performed in biological triplicates.

Article Snippet: For the ANXA1 augmentation, 2 mg mL−1 of Matrigel was supplemented with human recombinant ANXA1 (hrANXA1) (3770-AN-050, R&D Systems) to reach the final concentration of 0.74 μg mL−1 hrANXA1.

Techniques: Activity Assay, Staining