Journal: Journal of Advanced Research

Article Title: Novel function of macrophage migration inhibitory factor in regulating post-infarct inflammation and the therapeutic significance

doi: 10.1016/j.jare.2025.05.030

Figure Lengend Snippet: Macrophage migration inhibitory factor (MIF) promotes inflammatory cell migration and infiltration through C – C motif chemokine receptor 2 (CCR2) and C-X-C motif chemokine receptor (CXCR) 4 and promotes splenic monocyte mobilization via interaction with angiotensin Ⅱ type 1 receptor (AT-1R) after myocardial infarction (MI). A, peripheral blood mononuclear cells (PBMCs) chemotaxis in response to homogenized sham or infarct heart tissue from wild type (WT) mice at day 3 post-MI, or MI homogenate together with inhibition of C–C motif chemokine receptor 2 (CCR2), C-X-C motif chemokine receptor (CXCR) 2 and CXCR4, respectively. Cells/HPF, cells/high power field. PBMCs harvested from WT mice at 3 days post-MI. CCR2i, CCR2 inhibition. CXCR2i, CXCR2 inhibition. CXCR4i, CXCR4 inhibition. n = 4–8 per group. *** P < 0.001 vs. sham . B, PBMCs chemotaxis in response to recombinant human MIF (rMIF) or rMIF with inhibition of CCR2, CXCR2 and CXCR4, respectively. PBMCs harvested from WT mice at 3 days post-MI. n = 4–8 per group. *** P < 0.001 vs. control . C, Representative immunoblotting images for monocyte chemoattractant protein-1 (MCP-1), CXCR4 and internal reference protein (heat shock protein 60, HSP-60) in hearts from WT and global MIF deficient (MIFKO) mice with sham-operation or 72 h MI. D-E, Quantitative analysis of MCP-1 and CXCR4 expression. n = 6–9 per group. ** P < 0.01 vs. sham in the same genotype. *** P < 0.001 vs. sham in the same genotype. F, Splenic monocyte (from normal WT mice) chemotaxis in response to plasma from normal mice or mice with 1.5 h MI, or MI plasma together with losartan. Lor, losartan, AT-1R inhibitor. n = 6–8 per group. *** P < 0.001 vs. normal. G, Splenic monocytes (from normal WT mice) chemotaxis in response to rMIF and angiotensin II (Ang II), or together with losartan addition, respectively. n = 6–8 per group. ** P < 0.01 vs. Control. *** P < 0.001 vs. Control. H, Representative immunoblotting images for AT-1R and internal reference protein (HSP-60) in spleens from WT and global MIF deficient (MIFKO) mice with sham-operation and 1.5 h MI. I, Quantitative analysis of AT-1R expression. n = 8 per group. *** P < 0.001 vs. sham in the same genotype . J. Co-immunoprecipitation (Co-IP) assay using an anti-AT-1R antibody or IgG demonstrated the interaction between AT-1R and MIF, as well as between AT-1R and CD74 in splenic monocytes and spleen tissue. Input, input sample. IgG, negative control sample. IP, Co-IP sample.

Article Snippet: For both groups 1 and 2, PBMCs were pre-treated for 1 h using DMEM with or without CCR2 inhibitor (CCR2 antagonist 4 hydrochloride, 100 mM), CXCR2 inhibitor (SB225002, 800 nM) or CXCR4 inhibitor (Plerixafor, AMD3100, 10 μM) (all from MedChemExpress, USA).

Techniques: Migration, Chemotaxis Assay, Inhibition, Recombinant, Control, Western Blot, Expressing, Clinical Proteomics, Co-Immunoprecipitation Assay, Negative Control

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: ATG16L1 Regulates Reparative Function of Peritoneal Macrophages During Acute Drug-induced Liver Injury

doi: 10.1016/j.jcmgh.2025.101674

Figure Lengend Snippet: ATG16L1 in PMs promotes hepatocyte proliferation via the IL-10-CXCR2 axis. PMs were injected into APAP-treated WT recipient mice. Liver tissues were collected at 24 and 48 hours post DILI. IHC for PCNA ( A ). WB ( B ) and qPCR ( C, D ) for CXCR2 and PCNA expression in liver tissues at 48 hours post DILI. Primary hepatocytes were co-cultured with PMs stimulated with HA. qPCR for CXCR2 and PCNA in hepatocytes ( E, F ). PMs were stimulated by HA combined with NAC or DC-LC3in-D5. IL-10 levels were assessed by qPCR and ELISA ( G–J ). IL-10 neutralizing antibody was added into the co-cultured primary hepatocytes and PMs stimulated with HA. Expression of CXCR2 and PCNA in hepatocytes ( K–M ). (n = 6/group). Data are presented as mean ± SEM. Each point represents an independent experiment. ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001, ∗∗∗∗ P < .0001.

Article Snippet: Rabbit anti-mouse ATG16L1 (1:1000, ab187671, Abcam), rabbit anti-mouse CD44 (1:1000, ab243894, Abcam), rabbit anti-mouse OAS3 (1:1000, 21915-1-AP, proteintech), rabbit anti-mouse SLFN5 (1:1000, AF15102, AIFang biological), rabbit anti-mouse DHX58 (1:1000, 11355-1-AP, proteintech), rabbit anti-mouse EIF2AK2 (1:5000, 18244-1-AP, proteintech), rabbit anti-mouse TRIM21 (1:5000, 12108-1-AP, proteintech), rabbit anti-mouse MerTK (1:1000, 27900-1-AP, proteintech), rabbit anti-mouse Axl (1:1000, ab215205, Abcam), rabbit anti-mouse TYRO3 (1:1000, 28513-1-AP, proteintech), rabbit anti-mouse TIM4 (1:1000, ab47637, Abcam), rabbit anti-mouse CXCR2 (1:1000, 20634-1-AP, proteintech), rabbit anti-mouse LC3 (1:1000, ab192890, Abcam), rabbit anti-mouse p62 (1:1000, ab109012, Abcam), rabbit anti-mouse PCNA (1:1000, ab29, Abcam), and mouse anti-mouse β-actin (1:1000, #3700S, Cell Signaling Technology) were used.

Techniques: Injection, Expressing, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: medRxiv

Article Title: Altered neutrophil G-protein receptor signalling linked to impaired chemotaxis and increased ROS and NET production in older people with frailty

doi: 10.64898/2025.12.16.25342352

Figure Lengend Snippet: (A) Expression of CXCR1 (CD181), CXCR2 (CD182) and CD177 via flow cytometry. (B) Removal of the RA group resulted in significant differences. (C) Chemotaxis of neutrophils towards IL-8, fMLP or random migration (UT). (D) ERK-mediated signalling network predicted to be regulating chemotaxis in FR neutrophils (adj. p=5.23×10 -9 ). Red = up-regulated gene expression, Orange = predicted activation. Representative western blots and densitometry (n=4) for (E) phosphorylated ERK (ERK-P) in untreated (UT) and 5 min IL-8 treated neutrophils and (F) phosphorylated RAC-1 (RAC1-P) in freshly isolated, 0h neutrophils. Purple = FR (n=4-10), Pink = RA (n=4-9), Orange = HO (n=4-10), Green = HY (n=4-7). Analysed by ANOVA (*p<0.05, **p<0.01, ***p<0.001).

Article Snippet: Antibodies used were CD177 FITC Monoclonal Antibody (MEM-166, Thermo Fisher), CD54 (ICAM-1) PE-Vio615, REAfinity antibody (Miltenyi), CD181 (CXCR1) FITC REAfinity antibody (Miltenyi), CD182 (CXCR2) APC, REAfinity antibody (Miltenyi), REAfinity isotype controls (APC, PE-Vio615, FITC, Miltenyi) or unstained (US) controls.

Techniques: Expressing, Flow Cytometry, Chemotaxis Assay, Migration, Gene Expression, Activation Assay, Western Blot, Isolation