Journal: Annals of Translational Medicine

Article Title: Serum amyloid A1 as a biomarker for radiation dose estimation and lethality prediction in irradiated mouse

doi: 10.21037/atm.2019.12.27

Figure Lengend Snippet: Primer sequences used for quantitative PCR

Article Snippet: Serum proteins such as SAA1 and procalcitonin (PCT) were measured using enzyme-linked immunosorbent assay (ELISA) kits (Mouse SAA1: EK1190, Boster, Wuhan, China; Human SAA1: EK1544, Boster, Wuhan, China; PCT: E10371m, CUSABIO, Wuhan, China) according to the manufacturer’s instructions.

Techniques:

Journal: Annals of Translational Medicine

Article Title: Serum amyloid A1 as a biomarker for radiation dose estimation and lethality prediction in irradiated mouse

doi: 10.21037/atm.2019.12.27

Figure Lengend Snippet: Radiation time response in mouse SAA1 measured using ELISA in 8 Gy irradiated female C57BL/6J mice at 0, 1, 2, 4, and 6 hours post-irradiation. n=3 per group. *, P<0.05 in the irradiated mice compared with control mice.

Article Snippet: Serum proteins such as SAA1 and procalcitonin (PCT) were measured using enzyme-linked immunosorbent assay (ELISA) kits (Mouse SAA1: EK1190, Boster, Wuhan, China; Human SAA1: EK1544, Boster, Wuhan, China; PCT: E10371m, CUSABIO, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Irradiation

Journal: Annals of Translational Medicine

Article Title: Serum amyloid A1 as a biomarker for radiation dose estimation and lethality prediction in irradiated mouse

doi: 10.21037/atm.2019.12.27

Figure Lengend Snippet: Time and dose response of SAA1 after TBI. (A) SAA1 was measured using ELISA in 0, 1, 2, 4, 8 and 12 Gy irradiated female C57BL/6J mice at 0.125, 0.5, 1, 2, 3, 5 and 7 days post-irradiation. SAA1 dose-dependent change at (B) 0.25, (C) 0.5, (D) 1, (E) 2, (F) 3, (G) 5 and (H) 7 days after exposure to radiation. Each black dot represents one animal. Error bars indicate ± 1 SD for each radiation exposure group. n=8 per group (n=7 at 7 days after 8 Gy, n=2 at 7 days after 12 Gy). SAA1, serum amyloid A1; TBI, total body irradiation.

Article Snippet: Serum proteins such as SAA1 and procalcitonin (PCT) were measured using enzyme-linked immunosorbent assay (ELISA) kits (Mouse SAA1: EK1190, Boster, Wuhan, China; Human SAA1: EK1544, Boster, Wuhan, China; PCT: E10371m, CUSABIO, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Irradiation

Journal: Annals of Translational Medicine

Article Title: Serum amyloid A1 as a biomarker for radiation dose estimation and lethality prediction in irradiated mouse

doi: 10.21037/atm.2019.12.27

Figure Lengend Snippet: P values associated with two-tailed Student’s t -test for SAA1 concentrations after comparing control and irradiated groups

Article Snippet: Serum proteins such as SAA1 and procalcitonin (PCT) were measured using enzyme-linked immunosorbent assay (ELISA) kits (Mouse SAA1: EK1190, Boster, Wuhan, China; Human SAA1: EK1544, Boster, Wuhan, China; PCT: E10371m, CUSABIO, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Irradiation

Journal: Annals of Translational Medicine

Article Title: Serum amyloid A1 as a biomarker for radiation dose estimation and lethality prediction in irradiated mouse

doi: 10.21037/atm.2019.12.27

Figure Lengend Snippet: SAA1 mRNA time-dependent increase after radiation in (A) liver, (B) lung, (C) thymus, (D) spleen, (E) bone marrow and (F) small intestine measured using Quantitative PCR in control and 8 Gy irradiated female C57BL/6J mice at 0.125, 0.5, 1, 2, 3, 5 and 7 days post-irradiation. Error bars indicate ± 1 SD for each radiation exposure group. n=6 to 8 per group. *, P<0.05, *, P<0.01, ***, P<0.001, and ****, P<0.0001 in the irradiated mice compared with the control mice. SAA1, serum amyloid A1.

Article Snippet: Serum proteins such as SAA1 and procalcitonin (PCT) were measured using enzyme-linked immunosorbent assay (ELISA) kits (Mouse SAA1: EK1190, Boster, Wuhan, China; Human SAA1: EK1544, Boster, Wuhan, China; PCT: E10371m, CUSABIO, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Real-time Polymerase Chain Reaction, Irradiation

Journal: Annals of Translational Medicine

Article Title: Serum amyloid A1 as a biomarker for radiation dose estimation and lethality prediction in irradiated mouse

doi: 10.21037/atm.2019.12.27

Figure Lengend Snippet: SAA1 expression and systemic infection parameters in the same mice. (A) SAA1 and LPS in serum and 16S rRNA in the liver of the same animals measured on 31 mice on day 0,1, 5 and 7 (7 mice on day 7, and 8 per group on day 0, 1 and 5) after exposure to 8 Gy TBI. (B) SAA1 and PCT in serum in the same animals was on 23 mice on days 0, 0.125, 0.5, 1, 2, 3, 5 and 7 (2 mice on days 7 and 3 per group on days 0, 0.125, 0.5, 1, 2, 3 and 5) after exposure to 12 Gy TBI. SAA1, serum amyloid A1; TBI, total body irradiation.

Article Snippet: Serum proteins such as SAA1 and procalcitonin (PCT) were measured using enzyme-linked immunosorbent assay (ELISA) kits (Mouse SAA1: EK1190, Boster, Wuhan, China; Human SAA1: EK1544, Boster, Wuhan, China; PCT: E10371m, CUSABIO, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Expressing, Infection, Irradiation

Journal: Annals of Translational Medicine

Article Title: Serum amyloid A1 as a biomarker for radiation dose estimation and lethality prediction in irradiated mouse

doi: 10.21037/atm.2019.12.27

Figure Lengend Snippet: Multiple linear regression analysis between serum SAA1 concentration and number of lymphocyte and irradiation dose

Article Snippet: Serum proteins such as SAA1 and procalcitonin (PCT) were measured using enzyme-linked immunosorbent assay (ELISA) kits (Mouse SAA1: EK1190, Boster, Wuhan, China; Human SAA1: EK1544, Boster, Wuhan, China; PCT: E10371m, CUSABIO, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Concentration Assay, Irradiation

Journal: Annals of Translational Medicine

Article Title: Serum amyloid A1 as a biomarker for radiation dose estimation and lethality prediction in irradiated mouse

doi: 10.21037/atm.2019.12.27

Figure Lengend Snippet: SAA1 cut-off values of classification within 2 days post-irradiation

Article Snippet: Serum proteins such as SAA1 and procalcitonin (PCT) were measured using enzyme-linked immunosorbent assay (ELISA) kits (Mouse SAA1: EK1190, Boster, Wuhan, China; Human SAA1: EK1544, Boster, Wuhan, China; PCT: E10371m, CUSABIO, Wuhan, China) according to the manufacturer’s instructions.

Techniques:

Journal: Annals of Translational Medicine

Article Title: Serum amyloid A1 as a biomarker for radiation dose estimation and lethality prediction in irradiated mouse

doi: 10.21037/atm.2019.12.27

Figure Lengend Snippet: PBI patterns and SAA1 response. (A) The liver of C57BL/6J mice was shielded in PBI-1 and PBI-3 and in the corresponding groups PBI-2 and PBI-4 the liver was exposed. (B) SAA1 concentration in serum and (C) mRNA expression in liver of control, total-body irradiation and PBI group at 12 hours after 8 Gy irradiation. Error bars indicate ±1 SD for each radiation exposure group. n=6 per group. **, P<0.01, ***, P<0.001, and ****, P<0.0001 in the irradiated mice compared with the control mice. PBI, partial body irradiation; SAA1, serum amyloid A1.

Article Snippet: Serum proteins such as SAA1 and procalcitonin (PCT) were measured using enzyme-linked immunosorbent assay (ELISA) kits (Mouse SAA1: EK1190, Boster, Wuhan, China; Human SAA1: EK1544, Boster, Wuhan, China; PCT: E10371m, CUSABIO, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Concentration Assay, Expressing, Irradiation

Journal: Annals of Translational Medicine

Article Title: Serum amyloid A1 as a biomarker for radiation dose estimation and lethality prediction in irradiated mouse

doi: 10.21037/atm.2019.12.27

Figure Lengend Snippet: Dynamic SAA1 concentration and corresponding date of death within 30 days after 10 Gy irradiation

Article Snippet: Serum proteins such as SAA1 and procalcitonin (PCT) were measured using enzyme-linked immunosorbent assay (ELISA) kits (Mouse SAA1: EK1190, Boster, Wuhan, China; Human SAA1: EK1544, Boster, Wuhan, China; PCT: E10371m, CUSABIO, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Concentration Assay, Irradiation

Journal: Annals of Translational Medicine

Article Title: Serum amyloid A1 as a biomarker for radiation dose estimation and lethality prediction in irradiated mouse

doi: 10.21037/atm.2019.12.27

Figure Lengend Snippet: Mean and standard deviation (SD) of SAA1 concentration in 92 healthy mice

Article Snippet: Serum proteins such as SAA1 and procalcitonin (PCT) were measured using enzyme-linked immunosorbent assay (ELISA) kits (Mouse SAA1: EK1190, Boster, Wuhan, China; Human SAA1: EK1544, Boster, Wuhan, China; PCT: E10371m, CUSABIO, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Standard Deviation, Concentration Assay

Journal: Annals of Translational Medicine

Article Title: Serum amyloid A1 as a biomarker for radiation dose estimation and lethality prediction in irradiated mouse

doi: 10.21037/atm.2019.12.27

Figure Lengend Snippet: Kaplan-Meier survival curves of mice. P value determined by log-rank test. Serum SAA1 in mice treated with amifostine before 10 Gy irradiation and in the 10 Gy group on day −4, 1, 3, 5 and 7. n=8 in 10 Gy + Amifostine group, n=20 in 10 Gy group. ***, P<0.001. SAA1, serum amyloid A1.

Article Snippet: Serum proteins such as SAA1 and procalcitonin (PCT) were measured using enzyme-linked immunosorbent assay (ELISA) kits (Mouse SAA1: EK1190, Boster, Wuhan, China; Human SAA1: EK1544, Boster, Wuhan, China; PCT: E10371m, CUSABIO, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Irradiation

Journal: Annals of Translational Medicine

Article Title: Serum amyloid A1 as a biomarker for radiation dose estimation and lethality prediction in irradiated mouse

doi: 10.21037/atm.2019.12.27

Figure Lengend Snippet: SAA1 concentration in the Amifostine group and corresponding death date within 30 days

Article Snippet: Serum proteins such as SAA1 and procalcitonin (PCT) were measured using enzyme-linked immunosorbent assay (ELISA) kits (Mouse SAA1: EK1190, Boster, Wuhan, China; Human SAA1: EK1544, Boster, Wuhan, China; PCT: E10371m, CUSABIO, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Concentration Assay, Irradiation

Journal: Annals of Translational Medicine

Article Title: Serum amyloid A1 as a biomarker for radiation dose estimation and lethality prediction in irradiated mouse

doi: 10.21037/atm.2019.12.27

Figure Lengend Snippet: Clinical parameters and corresponding SAA1 concentration in all 17 NPC patients

Article Snippet: Serum proteins such as SAA1 and procalcitonin (PCT) were measured using enzyme-linked immunosorbent assay (ELISA) kits (Mouse SAA1: EK1190, Boster, Wuhan, China; Human SAA1: EK1544, Boster, Wuhan, China; PCT: E10371m, CUSABIO, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Concentration Assay

Journal: Annals of Translational Medicine

Article Title: Serum amyloid A1 as a biomarker for radiation dose estimation and lethality prediction in irradiated mouse

doi: 10.21037/atm.2019.12.27

Figure Lengend Snippet: Effect of radiotherapy on serum SAA1 in nasopharyngeal carcinoma patients. (A) The scatter plot shows SAA1 concentration before radiotherapy and corresponding expression after radiotherapy in 17 patients with nasopharyngeal carcinoma (****, P<0.0001). (B) ROC curve of SAA1 as a biomarker for predicting radiation exposure in patients with nasopharyngeal carcinoma. SAA1, serum amyloid A1.

Article Snippet: Serum proteins such as SAA1 and procalcitonin (PCT) were measured using enzyme-linked immunosorbent assay (ELISA) kits (Mouse SAA1: EK1190, Boster, Wuhan, China; Human SAA1: EK1544, Boster, Wuhan, China; PCT: E10371m, CUSABIO, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Concentration Assay, Expressing, Biomarker Assay

Journal: Stroke and vascular neurology

Article Title: Extracellular vesicles bearing serum amyloid A1 exacerbate neuroinflammation after intracerebral haemorrhage.

doi: 10.1136/svn-2024-003525

Figure Lengend Snippet: Figure 3 Human serum amyloid A1 (SAA1) protein levels in plasma and extracellular vesicles correlated (EVs) with clinical and laboratory assessments. (A, B) Bar graph indicating the concentration of SAA1 protein in both EVs and plasma for the ICH group and the healthy controls group. Unit: ng/mL in EVs and μg/mL in plasma. **p<0.01, ***p<0.001 by the two-tailed unpaired Student’s t-test and data are presented as means±SEM. (A) n=6, 15 in the control and ICH group, (B) n=13, 19 in the control and the ICH group. (C, D) Linear dependence graph revealing the relationship between neutrophils or leucocytes and the SAA1 protein in EVs (n=15). Correlation was analysed by spearman correlation analysis and the dashed line indicates the 95% CI. (E–H) Correlation analysis between the EVs-derived, plasma-derived SAA1 levels and the clinical assessments, including NIHSS and haemorrhage volume, through a linear dependence graph (n=15). Correlation was analysed by spearman correlation analysis and the dashed line indicates the 95% CI. ICH, intracerebral haemorrhage; NIHSS, National Institutes of Health Stroke Scale.

Article Snippet: We used anti- mouse SAA1 mAb (AF2948, NOVUS, USA) to block SAA1 in vivo at a dose of 0.25 μg/g, as previously reported.24 To establish a control, we administered Goat IgG Isotype Control (AB- 108- C, NOVUS, USA).

Techniques: Clinical Proteomics, Concentration Assay, Two Tailed Test, Control, Derivative Assay

Journal: Stroke and vascular neurology

Article Title: Extracellular vesicles bearing serum amyloid A1 exacerbate neuroinflammation after intracerebral haemorrhage.

doi: 10.1136/svn-2024-003525

Figure Lengend Snippet: Figure 4 Exogenous SAA1 upregulates the cell counts of microglia and astrocytes. (A) Schematic representation of exogenous SAA1 inserting into the basal ganglia region at varying dosages (750 ng, 250 ng, 83 ng and 0 ng). Images of immunostaining of microglia and astrocyte activation on day three are provided. (B, D) Images of immunostaining (B) and quantification (D) of microglia (stained by a specific marker, Iba1) around the ipsilateral basal ganglia region at different doses of exogenous SAA1. Microglia counts per mm2: 9.3±2.2, 41.5±3.9, 64.0±4.0, 88.3±8.8 for 0 ng, 83 ng, 250 ng, and 750 ng, respectively. (C, E) Images of immunostaining of GFAP (glial fibrillar acidic protein, known as the astrocyte active marker) representing astrocytes (C) and quantification of astrocytes (E) at different doses of exogenous SAA1 in the ipsilateral basal ganglia region. Astrocyte counts per mm2: 36.0±3.8, 93.5±6.1, 134.0±3.5, 176.5±11.6 for 0 ng, 83 ng, 250 ng, and 750 ng, respectively. Scale bars=100 µm, inset scale bars=20 µm, n=4 in each group. ***p<0.001 by the Kruskal-Wallis test. Data are presented as means±SEM.

Article Snippet: We used anti- mouse SAA1 mAb (AF2948, NOVUS, USA) to block SAA1 in vivo at a dose of 0.25 μg/g, as previously reported.24 To establish a control, we administered Goat IgG Isotype Control (AB- 108- C, NOVUS, USA).

Techniques: Immunostaining, Activation Assay, Staining, Marker

Journal: Stroke and vascular neurology

Article Title: Extracellular vesicles bearing serum amyloid A1 exacerbate neuroinflammation after intracerebral haemorrhage.

doi: 10.1136/svn-2024-003525

Figure Lengend Snippet: Figure 5 Blocking SAA1 promotes microglia reactivity and leucocyte infiltration. (A) Bar graph indicating the elevation of plasma SAA1 levels in ICH mice as compared with the sham group. n= 5 mice in the ICH group and n=10 mice in the sham group. **p<0.01 by two-tailed unpaired Student’s t-test. (B) Schematic diagram depicting intracerebral haemorrhage induction followed by intravenous administration of anti-SAA1 mAb or IgG 1 hour later. Immune cell populations were assessed using flow cytometry on days 1 and 3 post-ICH initiation. (C) Flow cytometry gating strategy depicting immune cell populations in mice brain treated with anti-SAA1 antibody or IgG on days 1 and day three post-ICH induction. The graph illustrates CD45high leucocytes, including CD3+CD19- T lymphocytes and its subtypes: CD4+CD8− T and CD4-CD8+ T lymphocytes, CD3−CD19+ B lymphocytes, CD11b+Ly6G+ neutrophils, and CD11b+F4/80+ macrophages. It also illustrates CD45int CD11b+microglia, including its subtypes: CD86+ microglia and CD206+ microglia. All gates were set using fluorescence-minus-one (FMO) controls. (D) Bar graph indicates the number of microglia and their subtypes in ICH mice with anti-SAA1 antibody or IgG treatment from days 1 to 3. (E) Bar graph shows major brain infiltrated leucocytes, involving CD8+ T lymphocytes, B lymphocytes, and neutrophils in ICH mice with anti-SAA1 antibody or IgG treatment from days 1 to 3. Int, intermediate. n=5, 6, 5 on day 1 and n=5, 13, 10 on day 3 for sham, IgG and mAb group. n=5, 9, 7 on day 3 for CD86+ microglia group in sham, IgG and mAb group specially. *p<0.05, **p<0.01, ***p<0.001 by one-way ANOVA and Tukey’s test. Data are presented as means±SEM. ANOVA, analysis of variance; ICH, intracerebral haemorrhage.

Article Snippet: We used anti- mouse SAA1 mAb (AF2948, NOVUS, USA) to block SAA1 in vivo at a dose of 0.25 μg/g, as previously reported.24 To establish a control, we administered Goat IgG Isotype Control (AB- 108- C, NOVUS, USA).

Techniques: Blocking Assay, Clinical Proteomics, Two Tailed Test, Flow Cytometry, Fluorescence

Journal: Stroke and vascular neurology

Article Title: Extracellular vesicles bearing serum amyloid A1 exacerbate neuroinflammation after intracerebral haemorrhage.

doi: 10.1136/svn-2024-003525

Figure Lengend Snippet: Figure 6 Anti-SAA1 mAb administration alleviates brain injury in mice with ICH. (A) Schematic diagram illustrating the ICH followed by intravenous injection of anti-SAA1 mAb or IgG 1 hour later. Subsequently, mice underwent MRI and neurological evaluations on days 1 and 3 after ICH induction. (B) Neurological scores of the sham group and the ICH group treated with anti- SAA1 mAb or IgG on day 1 and day 3. The modified Neurological Severity Score (mNSS) and rotarod test were used to measure the neurological deficit. n=6, 12, 7 for sham, IgG and mAb group. *p<0.05 by two-way ANOVA. (C) MRI of lesion volume (red) and perihaemorrhagic oedema (PHE) volume (yellow) on day 1 and day 3 post-ICH. (D, E) Quantification of lesion volume and PHE volume using the MRI. n=4, 6 for IgG and mAb group on day 1, n=5, 8 for IgG and mAb group on day 3. *p<0.05 by two-tailed unpaired Student’s t-test. Data are presented as means±SEM. ANOVA, analysis of variance; ICH, intracerebral haemorrhage.

Article Snippet: We used anti- mouse SAA1 mAb (AF2948, NOVUS, USA) to block SAA1 in vivo at a dose of 0.25 μg/g, as previously reported.24 To establish a control, we administered Goat IgG Isotype Control (AB- 108- C, NOVUS, USA).

Techniques: Injection, Modification, Two Tailed Test

Journal: Cell reports

Article Title: Toll-like receptor 2 orchestrates a tumor suppressor response in non-small cell lung cancer

doi: 10.1016/j.celrep.2022.111596

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit anti-SAA1 , Biorbyt , Cat #orb228668; RRID:AB_2924684.

Techniques: Plasmid Preparation, Polymer, Staining, Virus, Recombinant, Sequencing, Software

Journal: Stroke and Vascular Neurology

Article Title: Extracellular vesicles bearing serum amyloid A1 exacerbate neuroinflammation after intracerebral haemorrhage

doi: 10.1136/svn-2024-003525

Figure Lengend Snippet: Human serum amyloid A1 (SAA1) protein levels in plasma and extracellular vesicles correlated (EVs) with clinical and laboratory assessments. ( A, B ) Bar graph indicating the concentration of SAA1 protein in both EVs and plasma for the ICH group and the healthy controls group. Unit: ng/mL in EVs and μg/mL in plasma. **p<0.01, ***p<0.001 by the two-tailed unpaired Student’s t-test and data are presented as means±SEM. ( A ) n=6, 15 in the control and ICH group, ( B ) n=13, 19 in the control and the ICH group. ( C, D ) Linear dependence graph revealing the relationship between neutrophils or leucocytes and the SAA1 protein in EVs (n=15). Correlation was analysed by spearman correlation analysis and the dashed line indicates the 95% CI. ( E–H ) Correlation analysis between the EVs-derived, plasma-derived SAA1 levels and the clinical assessments, including NIHSS and haemorrhage volume, through a linear dependence graph (n=15). Correlation was analysed by spearman correlation analysis and the dashed line indicates the 95% CI. ICH, intracerebral haemorrhage; NIHSS, National Institutes of Health Stroke Scale.

Article Snippet: To evaluate the exogenous effect of SAA1 on the mouse brain, we administered varying doses (0 ng, 83 ng, 250 ng, 750 ng) of SAA1 protein (CSB-EP020656MO, CUSABIO, China) dissolved in 1 μL of saline via intracerebral injection.

Techniques: Clinical Proteomics, Concentration Assay, Two Tailed Test, Control, Derivative Assay

Journal: Stroke and Vascular Neurology

Article Title: Extracellular vesicles bearing serum amyloid A1 exacerbate neuroinflammation after intracerebral haemorrhage

doi: 10.1136/svn-2024-003525

Figure Lengend Snippet: Exogenous SAA1 upregulates the cell counts of microglia and astrocytes. ( A ) Schematic representation of exogenous SAA1 inserting into the basal ganglia region at varying dosages (750 ng, 250 ng, 83 ng and 0 ng). Images of immunostaining of microglia and astrocyte activation on day three are provided. ( B, D ) Images of immunostaining ( B ) and quantification ( D ) of microglia (stained by a specific marker, Iba1) around the ipsilateral basal ganglia region at different doses of exogenous SAA1. Microglia counts per mm 2 : 9.3±2.2, 41.5±3.9, 64.0±4.0, 88.3±8.8 for 0 ng, 83 ng, 250 ng, and 750 ng, respectively. ( C, E ) Images of immunostaining of GFAP (glial fibrillar acidic protein, known as the astrocyte active marker) representing astrocytes ( C ) and quantification of astrocytes ( E ) at different doses of exogenous SAA1 in the ipsilateral basal ganglia region. Astrocyte counts per mm 2 : 36.0±3.8, 93.5±6.1, 134.0±3.5, 176.5±11.6 for 0 ng, 83 ng, 250 ng, and 750 ng, respectively. Scale bars=100 µm, inset scale bars=20 µm, n=4 in each group. ***p<0.001 by the Kruskal-Wallis test. Data are presented as means±SEM.

Article Snippet: To evaluate the exogenous effect of SAA1 on the mouse brain, we administered varying doses (0 ng, 83 ng, 250 ng, 750 ng) of SAA1 protein (CSB-EP020656MO, CUSABIO, China) dissolved in 1 μL of saline via intracerebral injection.

Techniques: Immunostaining, Activation Assay, Staining, Marker

Journal: Stroke and Vascular Neurology

Article Title: Extracellular vesicles bearing serum amyloid A1 exacerbate neuroinflammation after intracerebral haemorrhage

doi: 10.1136/svn-2024-003525

Figure Lengend Snippet: Blocking SAA1 promotes microglia reactivity and leucocyte infiltration. ( A ) Bar graph indicating the elevation of plasma SAA1 levels in ICH mice as compared with the sham group. n= 5 mice in the ICH group and n=10 mice in the sham group. **p<0.01 by two-tailed unpaired Student’s t-test. ( B ) Schematic diagram depicting intracerebral haemorrhage induction followed by intravenous administration of anti-SAA1 mAb or IgG 1 hour later. Immune cell populations were assessed using flow cytometry on days 1 and 3 post-ICH initiation. ( C ) Flow cytometry gating strategy depicting immune cell populations in mice brain treated with anti-SAA1 antibody or IgG on days 1 and day three post-ICH induction. The graph illustrates CD45 high leucocytes, including CD3 + CD19 - T lymphocytes and its subtypes: CD4 + CD8 − T and CD4 - CD8 + T lymphocytes, CD3 − CD19 + B lymphocytes, CD11b + Ly6G + neutrophils, and CD11b + F4/80 + macrophages. It also illustrates CD45 int CD11b + microglia, including its subtypes: CD86 + microglia and CD206 + microglia. All gates were set using fluorescence-minus-one (FMO) controls. ( D ) Bar graph indicates the number of microglia and their subtypes in ICH mice with anti-SAA1 antibody or IgG treatment from days 1 to 3. ( E ) Bar graph shows major brain infiltrated leucocytes, involving CD8 + T lymphocytes, B lymphocytes, and neutrophils in ICH mice with anti-SAA1 antibody or IgG treatment from days 1 to 3. Int, intermediate. n=5, 6, 5 on day 1 and n=5, 13, 10 on day 3 for sham, IgG and mAb group. n=5, 9, 7 on day 3 for CD86 + microglia group in sham, IgG and mAb group specially. *p<0.05, **p<0.01, ***p<0.001 by one-way ANOVA and Tukey’s test. Data are presented as means±SEM. ANOVA, analysis of variance; ICH, intracerebral haemorrhage.

Article Snippet: To evaluate the exogenous effect of SAA1 on the mouse brain, we administered varying doses (0 ng, 83 ng, 250 ng, 750 ng) of SAA1 protein (CSB-EP020656MO, CUSABIO, China) dissolved in 1 μL of saline via intracerebral injection.

Techniques: Blocking Assay, Clinical Proteomics, Two Tailed Test, Flow Cytometry, Fluorescence

Journal: Stroke and Vascular Neurology

Article Title: Extracellular vesicles bearing serum amyloid A1 exacerbate neuroinflammation after intracerebral haemorrhage

doi: 10.1136/svn-2024-003525

Figure Lengend Snippet: Anti-SAA1 mAb administration alleviates brain injury in mice with ICH. ( A ) Schematic diagram illustrating the ICH followed by intravenous injection of anti-SAA1 mAb or IgG 1 hour later. Subsequently, mice underwent MRI and neurological evaluations on days 1 and 3 after ICH induction. ( B ) Neurological scores of the sham group and the ICH group treated with anti-SAA1 mAb or IgG on day 1 and day 3. The modified Neurological Severity Score (mNSS) and rotarod test were used to measure the neurological deficit. n=6, 12, 7 for sham, IgG and mAb group. *p<0.05 by two-way ANOVA. ( C ) MRI of lesion volume (red) and perihaemorrhagic oedema (PHE) volume (yellow) on day 1 and day 3 post-ICH. ( D, E ) Quantification of lesion volume and PHE volume using the MRI. n=4, 6 for IgG and mAb group on day 1, n=5, 8 for IgG and mAb group on day 3. *p<0.05 by two-tailed unpaired Student’s t-test. Data are presented as means±SEM. ANOVA, analysis of variance; ICH, intracerebral haemorrhage.

Article Snippet: To evaluate the exogenous effect of SAA1 on the mouse brain, we administered varying doses (0 ng, 83 ng, 250 ng, 750 ng) of SAA1 protein (CSB-EP020656MO, CUSABIO, China) dissolved in 1 μL of saline via intracerebral injection.

Techniques: Injection, Modification, Two Tailed Test

Journal: Animals : an Open Access Journal from MDPI

Article Title: Induction of Serum Amyloid A3 in Mouse Mammary Epithelial Cells Stimulated with Lipopolysaccharide and Lipoteichoic Acid

doi: 10.3390/ani11061548

Figure Lengend Snippet: Alignment of amino acid sequences of mouse serum amyloid A (SAA) isoforms. The sequences (AAEKISDGREAFOE and QRWVQFMKEAG) of synthetic SAA1 and SAA3 peptides for immunization to rabbits are underlined. Accession numbers are shown in parentheses.

Article Snippet: Subsequently, the membrane was incubated with the primary anti-mouse SAA1 rabbit polyclonal antibody (1:500, #PAA885Mu01, Cloud-Clone, Houston, TX, USA) or anti-SAA3 rat monoclonal antibody [JOR110A] (1:50, #ab231680, Abcam, Cambridge, UK) diluted with 1% nonfat dried skim milk in TBST for 1 h at room temperature.

Techniques:

Journal: Animals : an Open Access Journal from MDPI

Article Title: Induction of Serum Amyloid A3 in Mouse Mammary Epithelial Cells Stimulated with Lipopolysaccharide and Lipoteichoic Acid

doi: 10.3390/ani11061548

Figure Lengend Snippet: Oligonucleotide primers used in quantitative real-time PCR.

Article Snippet: Subsequently, the membrane was incubated with the primary anti-mouse SAA1 rabbit polyclonal antibody (1:500, #PAA885Mu01, Cloud-Clone, Houston, TX, USA) or anti-SAA3 rat monoclonal antibody [JOR110A] (1:50, #ab231680, Abcam, Cambridge, UK) diluted with 1% nonfat dried skim milk in TBST for 1 h at room temperature.

Techniques: Sequencing

Journal: Animals : an Open Access Journal from MDPI

Article Title: Induction of Serum Amyloid A3 in Mouse Mammary Epithelial Cells Stimulated with Lipopolysaccharide and Lipoteichoic Acid

doi: 10.3390/ani11061548

Figure Lengend Snippet: Comparison of Saa1 and Saa3 mRNA expression levels in NMuMG cells treated with lipopolysaccharide (LPS) and lipoteichoic acid (LTA). Data are presented as the mean ± standard deviation of four independent experiments. Asterisks indicate significant difference compared with control levels: ** p < 0.01. NMuMG, normal murine mammary gland.

Article Snippet: Subsequently, the membrane was incubated with the primary anti-mouse SAA1 rabbit polyclonal antibody (1:500, #PAA885Mu01, Cloud-Clone, Houston, TX, USA) or anti-SAA3 rat monoclonal antibody [JOR110A] (1:50, #ab231680, Abcam, Cambridge, UK) diluted with 1% nonfat dried skim milk in TBST for 1 h at room temperature.

Techniques: Expressing, Standard Deviation

Journal: Animals : an Open Access Journal from MDPI

Article Title: Induction of Serum Amyloid A3 in Mouse Mammary Epithelial Cells Stimulated with Lipopolysaccharide and Lipoteichoic Acid

doi: 10.3390/ani11061548

Figure Lengend Snippet: Comparison of SAA1 (A and B) and SAA3 (C and D) protein expression levels in NMuMG cells treated with lipopolysaccharide (LPS) or lipoteichoic acid (LTA) by immunofluorescence analysis (IFA). ( A , C ) Fields of view where fluorescence intensity was measured. Scale bar = 20 µm. ( B , D ) The relative SAA1 and SAA3 protein expression levels in NMuMG cells following stimulation with LPS or LTA were normalized to those in untreated, control cells. Data are presented as the mean fluorescence of five or more random locations with vertical bars representing standard deviation. Asterisks indicate significant difference compared with control levels: * p < 0.05, ** p < 0.01. NMuMG, normal murine mammary gland.

Article Snippet: Subsequently, the membrane was incubated with the primary anti-mouse SAA1 rabbit polyclonal antibody (1:500, #PAA885Mu01, Cloud-Clone, Houston, TX, USA) or anti-SAA3 rat monoclonal antibody [JOR110A] (1:50, #ab231680, Abcam, Cambridge, UK) diluted with 1% nonfat dried skim milk in TBST for 1 h at room temperature.

Techniques: Expressing, Immunofluorescence, Fluorescence, Standard Deviation

Journal: Animals : an Open Access Journal from MDPI

Article Title: Induction of Serum Amyloid A3 in Mouse Mammary Epithelial Cells Stimulated with Lipopolysaccharide and Lipoteichoic Acid

doi: 10.3390/ani11061548

Figure Lengend Snippet: Detection of SAA1 and SAA3 protein expression in NMuMG cell supernatants treated with lipopolysaccharide (LPS) or lipoteichoic acid (LTA) by Western blot. ( A ) Detection of SAA1 in the cell supernatant. Membranes were incubated with the primary anti-mouse SAA1 antibody (dilution of 1:500, 60 s exposure time). ( B ) Detection of SAA3 in the cell supernatant. Membranes were incubated with the primary anti-mouse SAA3 antibody (dilution of 1:50, 120 s exposure time). PC, positive control; 1, recombinant murine SAA1 (rSAA1); 3, rSAA3; NMuMG, normal murine mammary gland.

Article Snippet: Subsequently, the membrane was incubated with the primary anti-mouse SAA1 rabbit polyclonal antibody (1:500, #PAA885Mu01, Cloud-Clone, Houston, TX, USA) or anti-SAA3 rat monoclonal antibody [JOR110A] (1:50, #ab231680, Abcam, Cambridge, UK) diluted with 1% nonfat dried skim milk in TBST for 1 h at room temperature.

Techniques: Expressing, Western Blot, Incubation, Positive Control, Recombinant