bottom chamber contained serum free rpmi sdf 1 cxcl12  (R&D Systems)

Journal: Science Advances

Article Title: Fibroblast-like synoviocytes exacerbate autoimmune arthritis upon psychological stress in an IL-6–dependent manner

doi: 10.1126/sciadv.aec9257

Figure Lengend Snippet: ( A ) Cell-cell communication among clusters determined by CellChat. ( B ) GO term enrichment analysis in Fibroblast2. ( C ) Chemokine expression in each cluster. ( D ) CXCL1 and CXCL12 expression by IL-6/soluble IL-6Rα stimulation at protein levels in FLS in AIA. N = 5 from five independent experiments. P values were determined by Friedman test followed by Dunn’s test. ( E ) CXCR2 expression in human RA synovial samples (phs001457.v1.p1). ( F ) CXCL1 expression by IL-6/soluble IL-6Rα stimulation at protein levels in human RA FLS. N = 9 from four independent experiments. P values were determined by Friedman test followed by Dunn’s test. ( G ) The expression of chemokine receptor Cxcr2 in AIA. ( H ) The expression of CXCR2 in peripheral blood (PB) and synovial fluid (SF) neutrophils in RA ( GSE154474 and GSE116899 ). ( I to L ) The chemokine receptor antagonist experiment in the stress-induced AIA model. N = 9 to 10 from three independent experiments. (I) Ankle thickness after the intervention. (J) Ankle thickness at day 24. (K) Representative images of ankles. (L) Cumulative data of the histological score. P values were determined by unpaired t test.

Article Snippet: Mouse CXCL1 and CXCL12 levels of the culture supernatant were determined using a Mouse CXCL1 ELISA kit (Proteintech, Rosemont, IL) or a Mouse CXCL12 ELISA Kit (Proteintech).

Techniques: Analysis, Expressing

Journal: Nature Communications

Article Title: Neoadjuvant chemotherapy interferes with tertiary lymphoid structure formation in Epstein-Barr virus-associated gastric cancer

doi: 10.1038/s41467-026-77546-5

Figure Lengend Snippet: A UMAP embedding revealing six distinct subtypes of neutrophils identified in scRNA-seq samples ( n = 21). B KEGG pathway enrichments (fold enrichment) in different neutrophil subtypes ( n = 21). C Pseudotime cell trajectory of neutrophils ( n = 21) colored by subtypes (left) and dynamic expression of key chemokine receptors along the trajectory (right). D Dot plot showing expression levels and percentages of key chemokine receptors across the six neutrophil subtypes ( n = 21). E UMAP plot (left) showing re-clustering of neutrophils ( n = 21) into two functional states: aging neutrophils (Neu_aging) and inflammatory neutrophils (Neu_inflam). Feature plots (right) showing expression of aging markers SELL (CD62L, downregulated) and CXCR4 (upregulated). F Spatial feature maps ( n = 11), from top to bottom: spatial regions and regional TLSs; CXCR4-CXCL12 interaction scores; aging neutrophil abundance; and CXCR4 expression derived from aging neutrophils. Red arrows indicate co-localization of aging neutrophils at the periphery of IM-TLSs. G Bubble plot showing CXCL signaling communication probabilities between neutrophil subtypes and other microenvironmental populations in scRNAseq samples ( n = 21). Significance assessed by one-sided permutation test (100 permutations). H Representative mIHC images showing co-localization of aging neutrophils (CD66b + CXCR4+) and CXCL12-expressing stromal cells (α-SMA + CXCL12+) around a TLS (CD20 + B cells). Images are representative of 216 independent patient samples examined. Scale bar, 100 μm. I. Schematic of the in vitro Transwell assay evaluating CXCL12-mediated chemotaxis of aging neutrophils (Created in BioRender. Bi, J. (2026) https://BioRender.com/bhu0jn4 ). Representative FACS histograms of CXCR4 expression on neutrophils indirectly co-cultured with J EBV-positive SNU719 or K EBV-negative AGS cells, with or without 5-FU (NC vs. 5-FU). FACS gating strategy shown in Supplementary Fig. . Transwell migration assays ( n = 3) showing the chemotactic effect of CXCL12 on neutrophils co-cultured with L SNU719 or M AGS across treatment groups. FACS gating strategy shown in Supplementary Fig. . Two-sided Student’s t test with Bonferroni correction for multiple comparisons (ns, P ≥ 0.05; * P < 0.05; ** P < 0.01; *** P < 0.001). Exact P values are shown in .

Article Snippet: The markers incorporated in Panel 3 were PanCK (1:2000, Clone PDH09-10, Cat# HA601138 , HUABIO), BZLF1 (1:400, Clone BZ1, Cat#sc-53904, SANTA CRUZ), CD20 (1:2000, Clone #06, Cat#11007-MM06, Sino Biological), CD23 (1:500, Clone PD00-03, Cat#HA721139, HUABIO), CD66b (1:20,000, Clone EPR25354-2, Cat#ab300122, Abcam), MPO(1:200, Polyclonal, Cat#AF3667, R&D), CitH3(1:200, Polyclonal, Cat#ab5103, Abcam), CXCR4 (1:500, Clone UMB2, Cat#ab124824, Abcam), CXCL12 (1:100, Polyclonal, Cat#17402-1-AP, proteintech), αSMA (1:8000, Polyclonal, Cat#14395-1-AP, proteintech), and DAPI.

Techniques: Expressing, Functional Assay, Derivative Assay, In Vitro, Transwell Assay, Chemotaxis Assay, Cell Culture, Migration

Journal: Nature Communications

Article Title: Neoadjuvant chemotherapy interferes with tertiary lymphoid structure formation in Epstein-Barr virus-associated gastric cancer

doi: 10.1038/s41467-026-77546-5

Figure Lengend Snippet: Under the stressful conditions induced by 5-FU-based neoadjuvant chemotherapy, latent EBV undergoes lytic reactivation in cancer cells. Upon reactivation and cancer cell lysis, EBV releases viral pathogen-associated molecular patterns (PAMPs), including viral DNA, RNA, and proteins, which are subsequently recognized by pattern recognition receptors (PRRs), such as TLR7, on tumor-infiltrating neutrophils. This recognition triggers neutrophil phenotypic transformation toward an aging phenotype characterized by upregulated CXCR4 expression and enhanced NET-formation capacity. These aging neutrophils are recruited to the periphery of TLS via CXCR4-CXCL12 chemotaxis, where they release NETs that form a physical barrier isolating TLS as immunological “islands” This NET-mediated barrier prevents the continuous recruitment of antigen-presenting cells and disrupts TLS maturation and maintenance, ultimately contributing to poor prognosis in EBVaGC patients. (Created in BioRender. Bi, J. (2026) https://BioRender.com/3a9ekcx ).

Article Snippet: The markers incorporated in Panel 3 were PanCK (1:2000, Clone PDH09-10, Cat# HA601138 , HUABIO), BZLF1 (1:400, Clone BZ1, Cat#sc-53904, SANTA CRUZ), CD20 (1:2000, Clone #06, Cat#11007-MM06, Sino Biological), CD23 (1:500, Clone PD00-03, Cat#HA721139, HUABIO), CD66b (1:20,000, Clone EPR25354-2, Cat#ab300122, Abcam), MPO(1:200, Polyclonal, Cat#AF3667, R&D), CitH3(1:200, Polyclonal, Cat#ab5103, Abcam), CXCR4 (1:500, Clone UMB2, Cat#ab124824, Abcam), CXCL12 (1:100, Polyclonal, Cat#17402-1-AP, proteintech), αSMA (1:8000, Polyclonal, Cat#14395-1-AP, proteintech), and DAPI.

Techniques: Lysis, Transformation Assay, Expressing, Chemotaxis Assay