Journal: iScience

Article Title: Post-transplant cyclophosphamide prevents xenogeneic graft-versus-host disease while depleting proliferating regulatory T cells

doi: 10.1016/j.isci.2023.106085

Figure Lengend Snippet:

Article Snippet: Human HS Cytokine A Premixed Mag Luminex Performance Assay,Analyte: IL-2, TNF-alpha, IFN-gamma , , Bio-Techne Ltd , Cat# FCSTM09-03 , .

Techniques: Staining, Recombinant, Activation Assay, Lysis, Cell Isolation, Selection, Flow Cytometry, Luminex, Cell Stimulation, Sequencing, Isolation, Transduction, Luciferase, Expressing, Software, Imaging

Journal: International Journal of Biological Sciences

Article Title: CXCR4-dependent macrophage-to-fibroblast signaling contributes to cardiac diastolic dysfunction in heart failure with preserved ejection fraction

doi: 10.7150/ijbs.65802

Figure Lengend Snippet: CXCR4 governs a pro-inflammatory phenotype in macrophages. (A) QPCR analysis of CXCR4, IL-1β, IL-6, TNFα, and CCL5 in MΦ WT and MΦ MKO after HMGB1 treatment. (B) The KEGG pathway analysis of cluster in upregulated gene terms of MΦ WT and MΦ MKO after HMGB1 treatment. Top 10 of enriched KEGG term are listed. (C) QPCR analysis of PPARα, PPARβ, and PPARγ in MΦ WT and MΦ MKO with or without HMGB1 treatment. (D) Western blot analysis of CXCR4 and PPARγ in MΦ WT and MΦ MKO after HMGB1 treatment. (E) Immunofluorescence staining of PPARγ (red) (DAPI, blue) in MΦ WT and MΦ MKO after HMGB1 treatment. Scale bar = 50μm. (F) QPCR analysis of IL-1β, IL-6, TNFα, and CCL5 in MΦ WT and MΦ MKO with GW9662 or GW1929 after HMGB1 treatment. (G) Western blot analysis of pNF-κB p65 and NF-κB p65 in MΦ WT and MΦ MKO after HMGB1 treatment. (H) Western blot analysis of pNF-κB p65 and NF-κB p65 in MΦ WT and MΦ MKO with GW9662 or GW1929 after HMGB1 treatment. (I) Western blot analysis of pMEK, MEK, pERK1/2 and ERK1/2 in MΦ WT and MΦ MKO after HMGB1 treatment. (H) Western blot analysis of pNF-κB p65 and NF-κB p65 in MΦ WT and MΦ MKO withPD98095 after HMGB1 treatment. GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes; All data were analyzed using two-way ANOVA with Bonferroni's multiple comparisons test. *, p<0.05.

Article Snippet: Cells were stimulated with 1ug/ml HMGB1 (ab181949, Abcam, Cambridge, MA, USA) and 100ng/ml CXCL12 (ab270067, Abcam, Cambridge, MA, USA) for 12 h. Cells were harvested for protein extraction or RNA isolation with an RNA isolation kit.

Techniques: Western Blot, Immunofluorescence, Staining

Journal: International Journal of Biological Sciences

Article Title: CXCR4-dependent macrophage-to-fibroblast signaling contributes to cardiac diastolic dysfunction in heart failure with preserved ejection fraction

doi: 10.7150/ijbs.65802

Figure Lengend Snippet: CXCR4-deficient macrophages attenuate myofibroblast differentiation via CXCL3-CXCR2 axis. (A) Western blot analysis of FN and αSMA in CFs co-cultured in MΦ WT and MΦ MKO -CM with or without HMGB1 treatment. (B) QPCR analysis of FN and αSMA in CFs co-cultured in MΦ WT and MΦ MKO -CM with or without HMGB1 treatment. (C) Immunofluorescence staining of αSMA+ cells in CFs co-cultured in MΦ WT and MΦ MKO -CM with or without HMGB1 treatment. (αSMA. red; DAPI. blue), Scale bar = 50μm. (D) Volcano plot of RNA sequencing MΦ WT and MΦ MKO after HMGB1 treatment. (E) RNA array analysis of the indicated genes in MΦ WT and MΦ MKO after HMGB1 treatment. (F) QPCR and elisa analysis of CXCL3 MΦ WT and MΦ MKO -CM with or without HMGB1 treatment. (G) Western blot analysis of FN and αSMA in CFs co-cultured in MΦ WT -CM after HMGB1 treatment with or without CXCL3 Ab. (H) Western blot analysis of FN and αSMA in CFs co-cultured in MΦ MKO -CM after HMGB1 treatment with or without rmCXCL3 protein. (I) Western blot analysis of CXCR2 in CFs co-cultured in MΦ WT and MΦ MKO -CM with or without HMGB1 treatment. (J) Western blot analysis of CXCR2 in CFs co-cultured in MΦ WT -CM after HMGB1 treatment with or without CXCL3 Ab. (K) Western blot analysis of CXCR2 in CFs co-cultured in MΦ MKO -CM after HMGB1 treatment with or without rmCXCL3. (L) Western blot analysis of FN and αSMA in CFs co-cultured in MΦ WT -CM after HMGB1 treatment with or without SB225002. (M) Western blot analysis o FN and αSMA in CFs co-cultured in MΦ MKO -CM after HMGB1 treatment with or without rmCXCL3 or SB225002. FN, fibronectin; αSMA, a-smooth muscle actin; CFs, cardiac fibroblasts; MΦ WT macrophages derived from the bone marrow WT mice; MΦ MKO , macrophages derived from the bone marrow MKO mice. CM, conditioned medium. CXCL3, Chemokine (C‑X‑C) motif ligand 3. All data were analyzed using two-way ANOVA with Bonferroni's multiple comparisons test. *, p<0.05.

Article Snippet: Cells were stimulated with 1ug/ml HMGB1 (ab181949, Abcam, Cambridge, MA, USA) and 100ng/ml CXCL12 (ab270067, Abcam, Cambridge, MA, USA) for 12 h. Cells were harvested for protein extraction or RNA isolation with an RNA isolation kit.

Techniques: Western Blot, Cell Culture, Immunofluorescence, Staining, RNA Sequencing, Enzyme-linked Immunosorbent Assay, Derivative Assay