Journal: Journal of Neuroinflammation

Article Title: Single-cell RNA sequencing uncovers neutrophil clusters associated with autoimmune neuroinflammation

doi: 10.1186/s12974-026-03772-9

Figure Lengend Snippet: Enhanced SAA1 expression and NET formation in plasma of MS patients. (A) SAA1 protein expression was quantified by ELISA in plasma samples collected from MS patients and HC. (B) H3.1-nucleosome expression was quantified by ELISA in plasma samples collected from MS patients and HC

Article Snippet: The human SAA1 ELISA kit (DY3019-05; R & D Systems, Minneapolis, MN) detected human SAA1 in plasma samples from MS patients and HC.

Techniques: Expressing, Clinical Proteomics, Enzyme-linked Immunosorbent Assay

Journal: Journal of Advanced Research

Article Title: Characterization of immune features and discovery of potential biomarkers for ankylosing spondylitis using deep plasma proteomics

doi: 10.1016/j.jare.2025.05.052

Figure Lengend Snippet: Verification of potential biomarkers for AS. (A) Expression levels of SAA1, FERMT3, ILK, and TLN1 in plasma samples from active-stage AS patients, stable-stage AS patients, and healthy controls measured by proteomics. ROC curves for the individual protein (SAA1, FERMT3, ILK, and TLN1) as a predictor to classify AS patients from healthy controls (B), and active-stage AS patients from stable-stage AS patients (C). (D) ELISA analysis of SAA1, FERMT3, ILK, and TLN1 expressions in the plasma samples from an independent validation cohort of healthy controls (N, n = 24), active-stage AS (A, n = 27), and stable-stage AS patients (S, n = 28). Statistical analyses employed Wilcoxon rank-sum test for intergroup comparisons, with results reported as mean ± SEM. Significance thresholds based on BH-adjusted p -values were defined as: *, <0.05; **, <0.01; ***, <0.001.

Article Snippet: The human serum amyloid A (SAA1) ELISA kit (#OKIA00083) was purchased from Aviva Systems Biology Company (San Diego, CA, USA).

Techniques: Expressing, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Biomarker Discovery

Journal: Cell metabolism

Article Title: Cytoarchitectural multi-depot profiling reveals immune-metabolic crosstalk in human colon-associated adipose tissue

doi: 10.1016/j.cmet.2025.12.008

Figure Lengend Snippet: (A) Cell-cell communication analysis of SAA signaling pathway for the indicated depots and cells. Line colors and widths represent the sending cell and the strength of the signal, respectively. (B) Pseudobulk comparison of differentially expressed genes in epiploic vs. subcutaneous depots, highlighting the indicated signaling pathways. (C) Expression of SAA1 mRNA upon stimulation with LPS (10 ng/mL), TNF-α (2.5 ng/mL), IL-1β (10 ng/mL), or IL-6 (10 ng/mL) for 24 h. Data are presented as fold change (FC) over vehicle. Significant (<0.05) p values compared with the vehicle from one-way ANOVA are shown. (D) Relative expression of SAA1 (left) and SAA2 (right) mRNA in adipocytes, ASPCs, adipose-derived endothelial cells (adECs), and THP1-derived macrophages (THP1 M0). Significant (<0.05) p values compared with the cell-type-specific control from one-way ANOVA are shown. (E) Results from ELISA measurements of SAA in conditioned media from human adipocytes incubated without or with LPS for 72 h. Results are displayed as mean, error bars represent standard error of the mean, and statistical significance was calculated using Student’s t test. (F) Analyses of DNA methylation in the indicated cell classes for the promoter and gene body regions of SAA1 and SAA2 , respectively. Chromosomal localization and motifs for NFKB2/RELA/REL and STAT3 are indicated for both genes. (G) SAA1 and SAA2 mRNA expression in adipocytes treated with or without LPS in the presence or absence of different inhibitors described in the main text. Data are presented as FC over vehicle. Significant (<0.05) p values compared with the vehicle from one-way ANOVA are shown. For (C), (D), and (G), data are displayed as geometric mean ± 95% confidence intervals with technical replicates displayed from three independent experiments. ASPC, adipose stromal and progenitor cell.

Article Snippet: Human recombinant SAA1 protein , OriGene , Cat#TP310664.

Techniques: Comparison, Protein-Protein interactions, Expressing, Derivative Assay, Control, Enzyme-linked Immunosorbent Assay, Incubation, DNA Methylation Assay

Journal: Cell metabolism

Article Title: Cytoarchitectural multi-depot profiling reveals immune-metabolic crosstalk in human colon-associated adipose tissue

doi: 10.1016/j.cmet.2025.12.008

Figure Lengend Snippet: (A) Top Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways based on differentially expressed genes from comparisons of SAA1- or LPS-treated vs. control-treated adipose-derived stromal-vascular cells. (B) Network view of the six common SAA1- and LPS-regulated pathways showing shared and treatment-specific genes. (C) Expression scores for SAA1- (left) and LPS-responsive (right) genes following a 6-h incubation with either ligand in stromal-vascular cells from subcutaneous human WAT. Results are displayed on the uniform manifold approximation and projection panel from and violin plots for each indicated cell class. Note that adipocytes and mesothelial cells were removed as these are not present in the stromal-vascular fraction from the subcutaneous depot. (D) Representative immunostaining of epiploic adipose tissue using antibodies directed against markers for Adipo SAA (CES1) and macrophages (CD68). Hoechst33342 was used as the counterstain for nuclei. Scale bar, 100 μm. ASPC, adipose stromal and progenitor cell; NES, normalized enrichment score.

Article Snippet: Human recombinant SAA1 protein , OriGene , Cat#TP310664.

Techniques: Control, Derivative Assay, Expressing, Incubation, Immunostaining

Journal: NPJ Precision Oncology

Article Title: Comprehensive single-cell RNA analysis reveals intertumoral microenvironment heterogeneity and hub niche of carcinogenesis in thyroid cancer

doi: 10.1038/s41698-025-00924-7

Figure Lengend Snippet: A Circle plot summarizing the maximum number of interactions among individual cell types in metastatic lesions. The thickness of the connecting lines represents the interaction strength. B Comparison of the overall information flow, including the number and strength of interactions, within the inferred networks between PTC and metastatic lesions. C Heatmap displaying the potential outgoing and incoming interaction strength of each cellular cluster in PTC and metastatic lesions. D Heatmap illustrating the relative signaling contribution of each cell group based on the number and strength of interactions, comparing PTC with metastatic lesions. E Differential network centrality analysis based on EMT-like cancer-associated fibroblasts (CAFs), comparing PTC and metastatic lesions. F Bubble heatmap showing the cell-cell communication of selected ligand-receptor pairs between EMT-like CAFs and CD8 + PDCD1 + T cells. Dot size indicates the P -value, while color represents the communication probability. G Multiplex immunohistochemistry (mIHC) validation of the cross-talk between SAA1+ CAFs and T cells via the PPIA-BSG ligand-receptor interaction.

Article Snippet: We performed multiplex immunofluorescence staining using the following primary antibodies: THBS1 rabbit anti-human antibody (Affinity Biosciences; catalog no. DF6848), CD47 rabbit anti-human antibody (Affinity Biosciences; catalog no. DF6649), CD68 rabbit anti-human antibody (Affinity Biosciences; catalog no. DF7518), APOE rabbit anti-human antibody (Affinity Biosciences; catalog no. AF5178), CD3 rabbit anti-human antibody (Affinity Biosciences; catalog no. DF6848), THBS1 rabbit anti-human antibody (Affinity Biosciences; catalog no. DF6594), BSG rabbit anti-human antibody (Affinity Biosciences; catalog no. AF5221), COL3A1 rabbit anti-human antibody (Affinity Biosciences; catalog no. AF5457), PPIA rabbit anti-human antibody (Proteintech Group; catalog no. 10720-1-AP), and SAA1 rabbit anti-human antibody (Proteintech Group; catalog no. 16721-1-AP).

Techniques: Comparison, Multiplex Assay, Immunohistochemistry, Biomarker Discovery

Journal: iScience

Article Title: TGFβ signaling in cancer-associated fibroblasts drives a hepatic gp130-dependent pro-metastatic inflammatory program in colorectal cancer

doi: 10.1016/j.isci.2025.113696

Figure Lengend Snippet: TGFβ1 signaling in primary CRC and LM-CRC CAFs induces expression of IL-6 (A) Schematic overview of workflow for CAF isolation and TGFβ1 pathway-targeted qPCR array. (B) RNA expression of TGFβ1 target genes in two primary CRC CAFs (CAF1 and CAF2) and one LM-CRC CAF. qPCR values have been log transformed. (C) Numerical values depicting the seven TGFβ1 target genes that showed consistent upregulation. (D) Fold change RNA expression of these seven TGFβ1 target genes (yellow symbols). (E) IL6 RNA expression in primary CRC CAFs (CAF4 and CAF5) and the CCD-18Co colon fibroblast line after stimulation with TGFβ1 or medium control. N = 3 CCD-18Co, CAF4; N = 2 CAF5 independent biological experiments. ∗∗ p ≤ 0.01 determined by paired T test. (F) Protein expression of IL-6 in tissue lysates of normal colon and primary CRC (left panel, N = 50) or normal liver and LM-CRC (right panel, N = 50) as determined by ELISA. ∗∗∗∗ p ≤ 0.0001, ∗ p ≤ 0.05 determined by unpaired T test. (G) IL6 RNA expression in unstimulated normal colon fibroblasts ( N = 8) or primary CRC CAFs ( N = 10). (H) RNA expression of IL6 in the original CMS classified cohort (CMS1: N = 457; CMS2: N = 1183; CMS3: N = 409; CMS4: N = 773). ∗∗∗∗ p ≤ 0.0001 determined by one-way ANOVA with correction for multiple testing (Dunnett’s test).

Article Snippet: SAA1 concentrations in patient sera were measured using a commercially available kit (Human SAA1 DuoSet ELISA, R&D Systems) and a decrease of ≥0.3 OD450 value between post and pre-surgery sera was considered significant.

Techniques: Expressing, Isolation, RNA Expression, Transformation Assay, Control, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: TGFβ signaling in cancer-associated fibroblasts drives a hepatic gp130-dependent pro-metastatic inflammatory program in colorectal cancer

doi: 10.1016/j.isci.2025.113696

Figure Lengend Snippet: IL-6 signaling in CAFs is partly regulated through autocrine TGFβ1 signaling (A) RNA expression of IL6 in CAFs derived either from the primary tumor (CRC CAF, N = 18) or liver metastasis (LM-CRC CAF, N = 6). ∗∗∗ p ≤ 0.001 determined by unpaired T test. (B) RNA-seq data from GSE46824 dataset showing IL6 RNA expression in an independent cohort of primary CRC CAFs ( N = 14) and LM-CRC CAFs ( N = 11). ∗∗∗ p ≤ 0.001 determined by unpaired T test. (C) IL-6 protein expression in unstimulated primary CRC CAF ( N = 8) and LM-CRC CAF ( N = 6) supernatant determined by ELISA. ∗∗ p ≤ 0.01 determined by unpaired T test. (D and E) IL-6 protein expression in TGFβ1-stimulated (5 ng/mL) primary CRC CAFs ( N = 8) (D) and LM-CRC CAFs ( N = 6) (E). ∗ p ≤ 0.05 determined by unpaired T test. (F) Fold change IL-6 protein expression in supernatant from TGFβ1-stimulated vs. unstimulated primary CRC CAFs ( N = 8) and LM-CRC CAFs ( N = 6). ns, p > 0.05 determined by unpaired T test. (G) Percentage decrease in IL-6 protein expression in supernatant in primary CRC ( N = 5) and LM-CRC ( N = 5) CAFs after inhibition with SB431532 (Alk5 inhibitor). ns, p > 0.05 determined by unpaired T test. (H) RNA-seq data from GSE46824 dataset (2B) showing TGFβ1 RNA expression in an independent cohort of primary CRC CAFs ( N = 14) and LM-CRC CAFs ( N = 11). ∗ p ≤ 0.05 determined by unpaired T test. (I) TGFβ1 protein expression in unstimulated primary CRC CAF ( N = 5) and LM-CRC CAF ( N = 6) supernatant determined by ELISA. ∗ p ≤ 0.05 determined by unpaired T test.

Article Snippet: SAA1 concentrations in patient sera were measured using a commercially available kit (Human SAA1 DuoSet ELISA, R&D Systems) and a decrease of ≥0.3 OD450 value between post and pre-surgery sera was considered significant.

Techniques: RNA Expression, Derivative Assay, RNA Sequencing, Expressing, Enzyme-linked Immunosorbent Assay, Inhibition

Journal: iScience

Article Title: TGFβ signaling in cancer-associated fibroblasts drives a hepatic gp130-dependent pro-metastatic inflammatory program in colorectal cancer

doi: 10.1016/j.isci.2025.113696

Figure Lengend Snippet: TGFβ isoforms are elevated in CMS4 subtype CRC and are capable of inducing IL-6 expression (A–C) RNA expression of the three TGFβ isoforms in the original CMS cohort (CMS1: N = 457; CMS2: N = 1183; CMS3: N = 409; CMS4: N = 773). ∗∗∗∗ p ≤ 0.0001 determined by one-way ANOVA with correction for multiple testing (Dunnett’s test). (D) Protein expression of TGFβ isoforms in the supernatant of primary CRC and LM-CRC CAFs 48 h after the start of serum starvation as determined by ELISA. (E and F) Expression of IL-6 in supernatant of primary CRC CAFs ( N = 7) (E) or LM-CRC CAFs ( N = 6) (F) 48 h after start stimulation with either TGFβ2 or TGFβ3. ∗ p ≤ 0.05 determined by paired T test.

Article Snippet: SAA1 concentrations in patient sera were measured using a commercially available kit (Human SAA1 DuoSet ELISA, R&D Systems) and a decrease of ≥0.3 OD450 value between post and pre-surgery sera was considered significant.

Techniques: Expressing, RNA Expression, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: TGFβ signaling in cancer-associated fibroblasts drives a hepatic gp130-dependent pro-metastatic inflammatory program in colorectal cancer

doi: 10.1016/j.isci.2025.113696

Figure Lengend Snippet: TGFβ-mediated IL-6 production by CAFs induces the expression of neutrophil chemoattractants SAA1 and CXCL5 in hepatocytes (A) Schematic overview of ‘CAF priming of Huh-7 hepatocytes’, referring to Huh-7 cells that are exposed to TGFβ1-stimulated CRC CAF CM. (B) Western blot for pSTAT3 and total STAT3 in wildtype Huh-7 cells that were stimulated with CAF CM or TGFβ1 CAF CM from 3 different CRC CAF lines. (C) RNA expression of SAA1 and CXCL5 in wildtype Huh-7 cells stimulated with DMEM 0% FCS (Control), IL-6 (50 ng/mL), CAF CM, or TGFβ1 CAF CM. N = 2 or N = 3 for the Control group, due to the very low SAA1 expression; for the other conditions, N = 3.∗∗∗ p ≤ 0.001, ∗∗ p ≤ 0.01, ∗ p ≤ 0.05 determined by one-way ANOVA with correction for multiple testing (Dunnett’s test). (D and E) RNA expression of SAA1 and CXCL5 in wildtype Huh-7 cells after CAF priming with blockade of specific components of the IL-6 signaling pathway. Siltuximab (anti-IL-6, 2 μg/mL), tocilizumab (anti-IL6R, 8 μg/mL), α-gp130 (anti-gp130, 8 μg/mL), tofacitinib (anti-JAK1/3, 5 μM), or human IgG isotype control (Bio X Cell; 2 μg/mL) were added to Huh-7 cells along with TGFβ1-stimulated CAF CM for 15 min. Subsequently, these different stimulations were applied to Huh-7 cells for 10 min. N = 1 for all different conditions. ∗∗∗∗ p ≤ 0.0001, ∗∗∗ p ≤ 0.001 determined by one-way ANOVA with correction for multiple testing (Dunnett’s test).

Article Snippet: SAA1 concentrations in patient sera were measured using a commercially available kit (Human SAA1 DuoSet ELISA, R&D Systems) and a decrease of ≥0.3 OD450 value between post and pre-surgery sera was considered significant.

Techniques: Expressing, Western Blot, RNA Expression, Control

Journal: iScience

Article Title: TGFβ signaling in cancer-associated fibroblasts drives a hepatic gp130-dependent pro-metastatic inflammatory program in colorectal cancer

doi: 10.1016/j.isci.2025.113696

Figure Lengend Snippet: The induction of neutrophil chemoattractants in hepatocytes is fully gp130-dependent (A) Western blot for pSTAT3 and total STAT3 in CAF CM-primed Huh-7 cells in which gp-130 signaling is blocked using α-gp130 antibody. α-gp130, anti-gp130. β-actin is used as a loading control. (B) Western blot for pSTAT3 and total STAT3 in vector control, gp130 KO , or gp130 KO rescued Huh-7 cells (gp130 KO+Rescue ) after stimulation with DMEM 0% (Control) or TGFβ1 CAF CM. (C) RNA expression of SAA1 in gp130 KO and gp130 KO+Rescue Huh-7 cells after CAF priming ( N = 3 independent biological experiments). ∗∗ p ≤ 0.01 determined by unpaired T test. (D) IHC of pSTAT3 in Huh-7 cells harboring a vector control, gp130 KO , or gp130 KO+Rescue after stimulation with DMEM 0% or after CAF priming. Scale bar, 50 μm.

Article Snippet: SAA1 concentrations in patient sera were measured using a commercially available kit (Human SAA1 DuoSet ELISA, R&D Systems) and a decrease of ≥0.3 OD450 value between post and pre-surgery sera was considered significant.

Techniques: Western Blot, Control, Plasmid Preparation, RNA Expression

Journal: iScience

Article Title: TGFβ signaling in cancer-associated fibroblasts drives a hepatic gp130-dependent pro-metastatic inflammatory program in colorectal cancer

doi: 10.1016/j.isci.2025.113696

Figure Lengend Snippet: The IL-6 family of cytokine-JAK-STAT signaling axis is active in CMS4 CRC in vivo (A) Schematic overview of the KPN GEMM organoid experiment. (B) Gene Set Enrichment analysis shows the Enrichment Score (ES) for the IL6-JAK-STAT signaling hallmark in KPN tumor tissue ( N = 3) and wildtype normal colon ( N = 3). (C) Normalized counts of IL6, IL11, and LIF in KPN tumor tissue (KPN) and normal colon tissue (WT). ∗∗∗∗ p ≤ 0.0001, ∗∗ p ≤ 0.01 determined by unpaired T test. (D) Quantification of automated scoring of Ly6G staining in pre-metastatic livers of KPN transplanted mice ( N = 4) and WT livers ( N = 5). (E) Representative IHC of pSTAT3, Ly6G, and GFP in pre-metastatic and metastatic livers in the KPN GEMM. Scale bar, 50 μm. (F) Waterfall plot of the difference in serum SAA1 OD450 value before and after surgery in 16 patients with primary CRC without liver metastasis.

Article Snippet: SAA1 concentrations in patient sera were measured using a commercially available kit (Human SAA1 DuoSet ELISA, R&D Systems) and a decrease of ≥0.3 OD450 value between post and pre-surgery sera was considered significant.

Techniques: In Vivo, Staining

Journal: The Journal of Clinical Investigation

Article Title: SAA1/FPR2 signaling between keratinocytes and neutrophils sustains chronic inflammation in Sweet syndrome

doi: 10.1172/JCI193566

Figure Lengend Snippet: ( A ) Ligand-receptor analysis reveals keratinocyte- and neutrophil-specific interactions. Keratinocytes expressed SAA1 transcripts and neutrophils expressed the FPR2 receptor (red dot, right-most column). ( B ) Dot plot demonstrating predominantly cell-specific expression of SAA1 and FPR2 transcripts. The dot size reflects the percentage of cells expressing the gene, and the color illustrates the level of gene expression. ( C ) Representative immunofluorescence staining images and quantification from 5 diseased and 5 control samples, confirming the expression of SAA1 and FPR2 in keratinocytes and neutrophils, respectively. Scale bars: 100 μm. ( D ) Dot plot comparing keratinocyte SAA1 and the control gene DEFB1 in different inflammatory skin conditions. The dot size reflects the percentage of cells expressing the gene, and the color illustrates the level of gene expression. ( E ) SAA1 secretion measured by ELISA in healthy neutrophils or healthy neutrophils exposed to keratinocytes. ( F ) Antibodies blocking SAA1 and FPR2 restored long-lived neutrophil lifespan to WT neutrophil levels. ( G ) Recombinant human SAA1 increased neutrophil survival at 72 hours ( n = 3 independent donors). Data indicate the mean ± SEM. * P < 0.05, ** P < 0.01, and *** P < 0.001, by 2-tailed, unpaired Student’s t test ( E and G ) and 1-way ANOVA with individual comparisons ( F ).

Article Snippet: Freshly isolated neutrophils from healthy donors were treated with recombinant human SAA1 (SAA1-257H, Creative Biomart).

Techniques: Expressing, Gene Expression, Immunofluorescence, Staining, Control, Enzyme-linked Immunosorbent Assay, Blocking Assay, Recombinant