Journal: Oncotarget

Article Title: ANXA1 inhibits miRNA-196a in a negative feedback loop through NF-kB and c-Myc to reduce breast cancer proliferation

doi: 10.18632/oncotarget.8875

Figure Lengend Snippet: A. miR196a expression was quantified by qPCR in 5 breast cancer cell lines and 1 breast epithelial cell line MCF10A. Results are represented as a fold change in mRNA levels after normalizing to U6 and comparing to MCF10A. B. ANXA1 expression and miR196a expression were plotted and a linear correlation was derived. C. Predicted targets of miR196a were quantified using qPCR and normalized against the housekeeping gene. D. ANXA1 protein expression was analyzed using western blot in MCF7 cells transfected with empty vector (EV) or miR196a plasmids. E. The luciferase reporter containing wild-type or mutant ANXA1 3′-UTR was co-transfected into HEK293T cells with miR-196 plasmids and control. Results shown are mean ± SEM of three independent experiments. * P < 0.05 ** P < 0.01 vs the indicated controls.

Article Snippet: The membranes were blocked with 5% non-fat dry milk in 1 x Tris-buffered saline and Tween 20 (TBST) of 0.1% v/v and incubated with Annexin-A1, c-Myc, p65 primary antibody (Cell Signaling Technology Inc, Danvers, MA, USA) overnight at 4°C and monoclonal alpha-tubulin (Sigma-Aldrich Co, St. Louis, MO, USA) and room-temperature with secondary antibody.

Techniques: Expressing, Derivative Assay, Western Blot, Transfection, Plasmid Preparation, Luciferase, Mutagenesis, Control

Journal: Oncotarget

Article Title: ANXA1 inhibits miRNA-196a in a negative feedback loop through NF-kB and c-Myc to reduce breast cancer proliferation

doi: 10.18632/oncotarget.8875

Figure Lengend Snippet: A. MiR-196a expression in MCF7 cells and MCF7 cells overexpressing ANXA1 was quantified by qPCR. Results are represented as a fold change in mRNA levels after normalizing to U6. B. Primary miR-196a, precursor miR-196a and mature miR-196a expression levels were quantified by qPCR in MCF7 cells after overexpression or silencing of ANXA1. * P < 0.05. ** P < 0.01 vs relevant controls.

Article Snippet: The membranes were blocked with 5% non-fat dry milk in 1 x Tris-buffered saline and Tween 20 (TBST) of 0.1% v/v and incubated with Annexin-A1, c-Myc, p65 primary antibody (Cell Signaling Technology Inc, Danvers, MA, USA) overnight at 4°C and monoclonal alpha-tubulin (Sigma-Aldrich Co, St. Louis, MO, USA) and room-temperature with secondary antibody.

Techniques: Expressing, Over Expression

Journal: Oncotarget

Article Title: ANXA1 inhibits miRNA-196a in a negative feedback loop through NF-kB and c-Myc to reduce breast cancer proliferation

doi: 10.18632/oncotarget.8875

Figure Lengend Snippet: A. ANXA1 binding to the promoter of miR196a was analyzed using CHIP. B. - E. . MCF7 cells were transfected with c-myc or p65 or empty vector (EV) control plasmids, and the level of pri-miR-196a or mature miR196a was quantified by qPCR. * P < 0.05, ** P < 0.01 vs empty vector control. F. . Binding of c-Myc to the promoter of miR-196a was analyzed using CHIP.

Article Snippet: The membranes were blocked with 5% non-fat dry milk in 1 x Tris-buffered saline and Tween 20 (TBST) of 0.1% v/v and incubated with Annexin-A1, c-Myc, p65 primary antibody (Cell Signaling Technology Inc, Danvers, MA, USA) overnight at 4°C and monoclonal alpha-tubulin (Sigma-Aldrich Co, St. Louis, MO, USA) and room-temperature with secondary antibody.

Techniques: Binding Assay, Transfection, Plasmid Preparation, Control

Journal: Oncotarget

Article Title: ANXA1 inhibits miRNA-196a in a negative feedback loop through NF-kB and c-Myc to reduce breast cancer proliferation

doi: 10.18632/oncotarget.8875

Figure Lengend Snippet: A. MCF7 cells were transfected with control or ANXA1 siRNA and the levels of c-Myc or ANXA1 expression was determined using qPCR. B. MCF7 stably transfected EV or ANXA1-V5 cells were analyzed for levels of c-Myc or ANXA1 mRNA expression C. C-Myc promoter activity and D. c-Myc protein expression. E. MCF7 stably transfected EV or ANXA1-V5 cells were treated with the c-myc inhibitor 10058-f4 and the levels of primary miR196a was determined using qPCR. F. MCF7 stably transfected EV or ANXA1-V5 cells were analyzed for NFKB luciferase activity and ANXA1 expression. G. MCF7 stably transfected ANXA1-V5 cells were transfected with p65 siRNA and c-Myc activity and p65 expression, and H. primary miR196a expression was measured. * P < 0.05, ** P < 0.01 vs relevant controls.

Article Snippet: The membranes were blocked with 5% non-fat dry milk in 1 x Tris-buffered saline and Tween 20 (TBST) of 0.1% v/v and incubated with Annexin-A1, c-Myc, p65 primary antibody (Cell Signaling Technology Inc, Danvers, MA, USA) overnight at 4°C and monoclonal alpha-tubulin (Sigma-Aldrich Co, St. Louis, MO, USA) and room-temperature with secondary antibody.

Techniques: Transfection, Control, Expressing, Stable Transfection, Activity Assay, Luciferase

Journal: Oncotarget

Article Title: ANXA1 inhibits miRNA-196a in a negative feedback loop through NF-kB and c-Myc to reduce breast cancer proliferation

doi: 10.18632/oncotarget.8875

Figure Lengend Snippet: A. , B. miR-196a plasmids were transfected into MDA-MB-231 cells or MCF-7 cells at the indicated doses and proliferation was assessed. C. Anti-miR196a or negative control were transfected into MCF7 cells at the indicated doses and proliferation was assessed. D. , E. 100ng miR-196a plasmids were transfected into MDA-MB-231 cells or MCF-7 cells at the indicated time points and proliferation was assessed. F. Proliferation was measured in MCF7 stably transfected EV or ANXA1-V5 cells G. MCF7 cells were transfected with control or ANXA1 siRNA and proliferation was assessed. H. MCF7 stably transfected EV or ANXA1-V5 cells were transfected with plasmids to miR196a and ANXA1 expression I. and proliferation was measured at the indicated days after transfection. * P < 0.05, ** P < 0.01 vs relevant controls. # P < 0.05 ANOVA vs miR196a.

Article Snippet: The membranes were blocked with 5% non-fat dry milk in 1 x Tris-buffered saline and Tween 20 (TBST) of 0.1% v/v and incubated with Annexin-A1, c-Myc, p65 primary antibody (Cell Signaling Technology Inc, Danvers, MA, USA) overnight at 4°C and monoclonal alpha-tubulin (Sigma-Aldrich Co, St. Louis, MO, USA) and room-temperature with secondary antibody.

Techniques: Transfection, Negative Control, Stable Transfection, Control, Expressing

Journal: Oncotarget

Article Title: ANXA1 inhibits miRNA-196a in a negative feedback loop through NF-kB and c-Myc to reduce breast cancer proliferation

doi: 10.18632/oncotarget.8875

Figure Lengend Snippet: BALB/c nu/nu athymic mice were injected with control MDA-MB231-luc cells or cells stably overexpressing miR196a into the mammary fat pad. A. Tumor size was measured every week post injection and tumor volume was calculated by LxWxW/2. B. Tumor volumes from tumors excised at week 3 were calculated by LxWxW/2. ** P < 0.01 vs control group. C. Bioluminescence imaging for mice from control group and miRNA group at week 1,2,3 post-injection. D. Quantification of bioluminescence data in tumor samples. ** P < 0.01 vs control group. E. , F. The expression of miR-196a or ANXA1 was conducted by qPCR in tumors from control and miRNA groups. Results are represented as a fold change in mRNA levels in tumors normalized to U6 or GAPDH, respectively. *** P < 0.001 vs control group.

Article Snippet: The membranes were blocked with 5% non-fat dry milk in 1 x Tris-buffered saline and Tween 20 (TBST) of 0.1% v/v and incubated with Annexin-A1, c-Myc, p65 primary antibody (Cell Signaling Technology Inc, Danvers, MA, USA) overnight at 4°C and monoclonal alpha-tubulin (Sigma-Aldrich Co, St. Louis, MO, USA) and room-temperature with secondary antibody.

Techniques: Injection, Control, Stable Transfection, Imaging, Expressing

Journal: Journal of Proteome Research

Article Title: Effect of Tamoxifen on Proteome Expression during In Vitro Myogenesis in Murine Skeletal Muscle C 2 C 12 Cells

doi: 10.1021/acs.jproteome.3c00340

Figure Lengend Snippet: Dilutions of Primary Antibody

Article Snippet: annexin A1 , Monoclonal IgG1 (mouse) , ProteinTech, 66344-1 , 1:2500.

Techniques:

Journal: Frontiers in Immunology

Article Title: GPCRs overexpression and impaired fMLP-induced functions in neutrophils from chronic kidney disease patients

doi: 10.3389/fimmu.2024.1387566

Figure Lengend Snippet: CKD neutrophils overexpress intracellular AnxA1. Increased FPR2 expression in CKD neutrophils (A) . AnxA1 expression in neutrophils at intracellular (B) and membrane compartments (C) . AnxA1 secretion from neutrophils (D) and apoptotic neutrophils (E) cultured for 18 hours under serum starvation conditions. (Control n= 3–11 and CKD n= 3–36) **p<0.01; ***p<0.001 CKD vs Control. ns, Not significant.

Article Snippet: Patients’ samples were incubated with anti-AnxA1 goat antibody (1:100) (AF3770; R&D SYSTEM) and anti-myeloperoxidase rabbit antibody (1:50; Biodesign International, Inc., Maine, USA) overnight at 4°C, followed by incubation with anti-goat Alexa Fluor 467 and anti-rabbit Alexa Fluor 488 secondary antibodies (1:200; Invitrogen).

Techniques: Expressing, Membrane, Cell Culture, Control

Journal: Frontiers in Immunology

Article Title: GPCRs overexpression and impaired fMLP-induced functions in neutrophils from chronic kidney disease patients

doi: 10.3389/fimmu.2024.1387566

Figure Lengend Snippet: Infiltrate neutrophils in CKD kidney biopsies are AnxA + . H&E (A–E) and PAS staining (F–J) . Myeloperoxidase (MPO) and AnxA1 labeling (K–O) . Five CKD biopsies were selected and stained for morphological, fibrosis visualization, and immunofluorescence. Figure 5D – Insert: * immune cells infiltrated; MPO and AnxA1 labeling. The image is representative of at least three fields analyzed on each biopsy.

Article Snippet: Patients’ samples were incubated with anti-AnxA1 goat antibody (1:100) (AF3770; R&D SYSTEM) and anti-myeloperoxidase rabbit antibody (1:50; Biodesign International, Inc., Maine, USA) overnight at 4°C, followed by incubation with anti-goat Alexa Fluor 467 and anti-rabbit Alexa Fluor 488 secondary antibodies (1:200; Invitrogen).

Techniques: Staining, Labeling, Immunofluorescence

Journal: BioImpacts

Article Title: Annexin A1, calreticulin and high mobility group box 1 are elevated in secondary progressive multiple sclerosis: Does immunogenic cell death occur in multiple sclerosis?

doi: 10.34172/bi.30264

Figure Lengend Snippet: Fig. 1. ANXA1 level in CSF sample of control group and SPMS patient. Bar graph related to ANXA1 measurement using standard ELISA assay that represents means ± SD measured in each group. A comparison between the results from control group and SPMS patient was done using a t-test. ELISA experiments were repeated at least three times. ** indicates: P < 0.01.

Article Snippet: Human ANXA1 commercial ELISA Kit (Elabscience; E-EL-H5512) was used to determine the level of ANXA1.

Techniques: Control, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Frontiers in Immunology

Article Title: Annexin A1 downregulates in vitro IL-1 β production in L. braziliensis -infected cells

doi: 10.3389/fimmu.2025.1685264

Figure Lengend Snippet: Differentially Expressed Genes in Clinical Samples from Cutaneous Leishmaniasis Patients. (A) Heatmap showing clustering of gene expression for PYCARD, CASP1, NLRP3, IL-10, IL1B, ANXA1, and FPR2 in peripheral blood from HS (n = 14) and CL patients (n = 50). Rows represent genes and columns represent individuals. The color scale indicates relative expression levels, ranging from blue (low expression) to yellow (high expression). (B-H) Differences in gene expression between CL patients and HS for (B) PYCARD , (C) NLRP3 , (D) FPR2 , (E) IL10 , (F) ANXA1, (G) IL1B , and (H) CASP1. PYCARD, CASP1 , and ANXA1 showed significantly higher expression in CL patients compared to HS. (I) Pearson correlation matrix of PYCARD, NLRP3, CASP1, IL1B, FPR2 , and ANXA1 expression in peripheral blood from CL patients (n = 50). Positive correlations are shown in green and negative correlations in red. (J) Serum levels of ANXA1 and (K) IL-1 β in CL patients (n = 50) and HS (n = 14), determined by ELISA. Results are expressed as mean ± standard deviation. Statistical analysis was performed using the Mann–Whitney test. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: After incubation, the infected cells were cultured in the presence or absence of recombinant Annexin A1 (rAnnexin A1) (R& D Systems, Minneapolis, MN, USA) at concentrations of 1 μ g/ml, as well as WRW4 (Tocris Bioscience, Bristol, UK) (23 μ M), an FPR2 inhibitor, for 4 and 48 hours.

Techniques: Gene Expression, Expressing, Enzyme-linked Immunosorbent Assay, Standard Deviation, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Annexin A1 downregulates in vitro IL-1 β production in L. braziliensis -infected cells

doi: 10.3389/fimmu.2025.1685264

Figure Lengend Snippet: CL patients show higher expression of inflammasome-related genes in active lesions. (A) Heat map showing expression of PYCARD , ANXA1 , CASP1 , IL1B , FPR2 , and NLRP3 in lesion biopsies from CL patients (n = 21) and healthy skin samples (n = 7), clustered by similarity. Blue indicates low expression, yellow indicates high expression. (B-H) Gene expression differences between CL lesions and healthy skin for (B) PYCARD , (C) NLRP3 , (D) FPR2 , (E) IL10 , (F) ANXA1 , (G) IL1B , and (H) CASP1 . (I) Correlation matrix for gene expression in CL lesions (n = 21) showing positive correlations (green) and negative correlations (red). Results are expressed as mean ± standard deviation. Statistical analysis was performed using the Mann–Whitney U test for (B-H) and Pearson correlation for (I) . * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: After incubation, the infected cells were cultured in the presence or absence of recombinant Annexin A1 (rAnnexin A1) (R& D Systems, Minneapolis, MN, USA) at concentrations of 1 μ g/ml, as well as WRW4 (Tocris Bioscience, Bristol, UK) (23 μ M), an FPR2 inhibitor, for 4 and 48 hours.

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

Journal: Frontiers in Immunology

Article Title: Annexin A1 downregulates in vitro IL-1 β production in L. braziliensis -infected cells

doi: 10.3389/fimmu.2025.1685264

Figure Lengend Snippet: ANXA1 down-regulates IL-1 β Production in CL. Levels of (A) ANXA1, (B) IL-1 β , and (C) IL-10 determined by ELISA in supernatants from CL lesion biopsy cultures (n = 8) and healthy skin from HS (n = 5). (D) IL-1 β and (E) IL-10 in lesion biopsy cultures (48 hours) from CL patients treated or not with rANXA1 (1 µ g). Results are expressed as mean ± standard deviation. Statistical analysis was performed using the Wilcoxon test. ** P < 0.01.

Article Snippet: After incubation, the infected cells were cultured in the presence or absence of recombinant Annexin A1 (rAnnexin A1) (R& D Systems, Minneapolis, MN, USA) at concentrations of 1 μ g/ml, as well as WRW4 (Tocris Bioscience, Bristol, UK) (23 μ M), an FPR2 inhibitor, for 4 and 48 hours.

Techniques: Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Frontiers in Immunology

Article Title: Annexin A1 downregulates in vitro IL-1 β production in L. braziliensis -infected cells

doi: 10.3389/fimmu.2025.1685264

Figure Lengend Snippet: ANXA1 is produced by L. braziliensis -infected macrophages. Monocytes-derived macrophages were infected with L. braziliensis (5:1) for 4 and 48 hours. (A) ANXA1, (B) IL-1 β and (C) IL-10 levels determined by ELISA. Results are expressed as mean ± standard deviation. Statistical analysis was performed using the Wilcoxon test. * P < 0.05, ** P < 0.01.

Article Snippet: After incubation, the infected cells were cultured in the presence or absence of recombinant Annexin A1 (rAnnexin A1) (R& D Systems, Minneapolis, MN, USA) at concentrations of 1 μ g/ml, as well as WRW4 (Tocris Bioscience, Bristol, UK) (23 μ M), an FPR2 inhibitor, for 4 and 48 hours.

Techniques: Produced, Infection, Derivative Assay, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Frontiers in Immunology

Article Title: Annexin A1 downregulates in vitro IL-1 β production in L. braziliensis -infected cells

doi: 10.3389/fimmu.2025.1685264

Figure Lengend Snippet: ANXA1 downregulates IL-1 β through the receptor FPR2 in L. braziliensis -infected macrophages. Levels of ANXA1, IL-1 β , and IL-10 in culture supernatants of monocyte-derived macrophages from healthy subjects (n = 9) infected with Leishmania braziliensis (5:1) in presence and absence of recombinant Annexin A1(1 µ g/mL) and WRW4(23 µ M/mL) at 4 and 48 hours. (A) ANXA1 levels, (B) IL-1 β levels, and (C) IL-10 in culture supernatants were determined by ELISA. Results are expressed as mean ± standard deviation. Statistical analysis was performed using the Wilcoxon test. * P < 0.05, ** P < 0.01.

Article Snippet: After incubation, the infected cells were cultured in the presence or absence of recombinant Annexin A1 (rAnnexin A1) (R& D Systems, Minneapolis, MN, USA) at concentrations of 1 μ g/ml, as well as WRW4 (Tocris Bioscience, Bristol, UK) (23 μ M), an FPR2 inhibitor, for 4 and 48 hours.

Techniques: Infection, Derivative Assay, Recombinant, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Frontiers in Immunology

Article Title: Annexin A1 downregulates in vitro IL-1 β production in L. braziliensis -infected cells

doi: 10.3389/fimmu.2025.1685264

Figure Lengend Snippet: ANXA1 does not affect phagocytic ability and parasite killing by macrophages infected with L. braziliensis . Effect of WRW4 and rANXA1 treatment on monocyte-derived macrophages from healthy subjects (n = 9) infected with Leishmania braziliensis (5:1) at 4 and 48 hours. rate infection (A) and the number of amastigotes per 100 cells (B) . Results are expressed as mean ± standard deviation. Statistical analysis was performed using the unpaired t-test. * P < 0.05, ** P < 0.01.

Article Snippet: After incubation, the infected cells were cultured in the presence or absence of recombinant Annexin A1 (rAnnexin A1) (R& D Systems, Minneapolis, MN, USA) at concentrations of 1 μ g/ml, as well as WRW4 (Tocris Bioscience, Bristol, UK) (23 μ M), an FPR2 inhibitor, for 4 and 48 hours.

Techniques: Infection, Derivative Assay, Standard Deviation