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Impaired chemotaxis in frailty neutrophils. (a) Expression of CXCR1 (CD181), <t>CXCR2</t> <t>(CD182)</t> and CD177 via flow cytometry. (b) A sub-group analysis of FR, HO and HY 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. (e) Representative western blot and densitometry (n = 4) for phosphorylated ERK (ERK-P) in untreated (UT) and 5 min IL-8 treated neutrophils. ERK-P intensity was normalised to Actin. 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).
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Impaired chemotaxis in frailty neutrophils. (a) Expression of CXCR1 (CD181), <t>CXCR2</t> <t>(CD182)</t> and CD177 via flow cytometry. (b) A sub-group analysis of FR, HO and HY 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. (e) Representative western blot and densitometry (n = 4) for phosphorylated ERK (ERK-P) in untreated (UT) and 5 min IL-8 treated neutrophils. ERK-P intensity was normalised to Actin. 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).
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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.

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

Impaired chemotaxis in frailty neutrophils. (a) Expression of CXCR1 (CD181), CXCR2 (CD182) and CD177 via flow cytometry. (b) A sub-group analysis of FR, HO and HY 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. (e) Representative western blot and densitometry (n = 4) for phosphorylated ERK (ERK-P) in untreated (UT) and 5 min IL-8 treated neutrophils. ERK-P intensity was normalised to Actin. 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).

Journal: Clinical and Experimental Immunology

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

doi: 10.1093/cei/uxag019

Figure Lengend Snippet: Impaired chemotaxis in frailty neutrophils. (a) Expression of CXCR1 (CD181), CXCR2 (CD182) and CD177 via flow cytometry. (b) A sub-group analysis of FR, HO and HY 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. (e) Representative western blot and densitometry (n = 4) for phosphorylated ERK (ERK-P) in untreated (UT) and 5 min IL-8 treated neutrophils. ERK-P intensity was normalised to Actin. 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, n = 5 per group), CD54 (ICAM-1, n = 7–10 per group) PE-Vio615, REAfinity antibody (Miltenyi, n = 7–10 per group), CD181 (CXCR1) FITC REAfinity antibody (Miltenyi, n = 7–10 per group), CD182 (CXCR2) APC, REAfinity antibody (Miltenyi, n = 7–10 per group), REAfinity isotype controls (APC, PE-Vio615, FITC, Miltenyi), or unstained (US) controls.

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