rabbit anti mouse cxcr2  (Proteintech)

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: Frontiers in Immunology

Article Title: Role of CXCL5 in Regulating Chemotaxis of Innate and Adaptive Leukocytes in Infected Lungs Upon Pulmonary Influenza Infection

doi: 10.3389/fimmu.2021.785457

Figure Lengend Snippet: CXCL5 regulated CXCL13 expression in pulmonary CD64+ macrophages/monocytes via the CXCL1/2/5-CXCR2 signaling pathway. (A–C) The protein levels of CXCL1/2 (A) , CXCL12 (B) , and CXCL13 (C) in the BALF of infected mice were measured by ELISA, and the level of CXCL5 was used as a reference (n=5). (D) RT-qPCR for IFN-α and IFN-β mRNA was performed in lung homogenates from WT and CXCL5 -/- mice before and after (3 d.p.i.) influenza infection. The fold changes in the mRNA expression levels of these genes were calculated using the 2 -ΔΔCt method of relative quantification with GAPDH as the endogenous reference gene. The relative fold changes in IFN-α and IFN-β expression in infected mice compared to normal mice are presented (n=4). (E, F) Percentages and numbers of CXCL13-expressing cells among single lung cells from WT and CXCL5 -/- mice at 3 d.p.i. (E) and 8 d.p.i. (F) as assessed by intracellular staining of CXCL13 protein (APC-CXCL13) and flow cytometry (n=4). (G, H) Percentages and numbers of CXCL13-expressing cells among pulmonary CD64+ cells from WT and CXCL5 -/- mice at 3 d.p.i. (G) and 8 d.p.i. (H) as assessed by intracellular staining of CXCL13 protein (APC-CXCL13) and flow cytometry (n=4). (I) Percentages and numbers of CXCL13-expressing cells among pulmonary CD11c+ cells from WT and CXCL5 -/- mice at 8 d.p.i. as assessed by intracellular staining of CXCL13 protein (APC-CXCL13) and flow cytometry (n=4). (J–L) Percentages and numbers of CD206 (J) , CD44 (K) , and CD274 (L) surface markers among pulmonary CD64+ cells from WT and CXCL5-/- mice at 3 d.p.i. (n=4). (M) Detection of the expression of mouse CXCR2 on total AMs and CD64+ AMs by western blotting. (N) RT-qPCR was performed on cell lysates of cultured CD64+ macrophages after 10 h of influenza challenge or unstimulated (control) conditions, and the fold changes were calculated as described in the MATERIALS AND METHODS section (n=4). (O) Expression levels of the indicated chemokines from supernatants of cultured CD64+ macrophages after 20 h of influenza challenge or unstimulated (control) conditions as determined by ELISA (n=4). (P) ELISA was used to determine the expression levels of CXCL13 protein in supernatants of cultured CD64+ macrophages after 20 h of influenza challenge and treatment with the indicated recombinant chemokine proteins, the CXCR2 antagonist SB225002, the PI3K inhibitor LY294002, and the MEK inhibitor PD98059 (n=3). The error bars represent the SDs. * P < 0.05 based on Student’s t -test.

Article Snippet: The membranes were stained with a rabbit anti-mouse CXCR2 polyclonal antibody (ab65968, Abcam, UK) and a goat anti-rabbit IgG H&L (HRP) polyclonal antibody (ab6721, Abcam, UK).

Techniques: Expressing, Infection, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Quantitative Proteomics, Staining, Flow Cytometry, Western Blot, Cell Culture, Control, Recombinant

Journal: Frontiers in Immunology

Article Title: Role of CXCL5 in Regulating Chemotaxis of Innate and Adaptive Leukocytes in Infected Lungs Upon Pulmonary Influenza Infection

doi: 10.3389/fimmu.2021.785457

Figure Lengend Snippet: CXCL5-CXCR2 signaling regulated pulmonary neutrophil and B cell recruitment and CXCL13 production from lung macrophages upon H1N1 infection. (A–E) WT mice were infected with H1N1 virus (1000 CCID 50 ) and treated with either CXCR2 antagonist or PBS as a control during 1-3 d.p.i. Microscopic view of BALF leukocytes from the infected mice collected at 3 d.p.i. and cytospun (×400 magnification) (A) . The protein level of CXCL13 in the BALF of the infected mice was measured at 3 d.p.i. by ELISA (n=4) (B) . Percentages and numbers of CXCL13-expressing cells among pulmonary CD64+ cells (C) , CD19+ B cells (D) , and CD44+/CD274+ CD64+ cells (E) from the infected mice at 3 d.p.i. by flow cytometry and counting (n=4). (F–J) WT mice were infected with H1N1 virus (1000 CCID 50 ) and orotracheally instilled with either CXCL5 recombinant protein or PBS as a control during 5-8 d.p.i. Microscopic view of BALF leukocytes from the infected mice collected at 8 d.p.i. and cytospun (×400 magnification) (F) . The protein level of CXCL13 in the BALF of the infected mice was measured at 8 d.p.i. by ELISA (n=4) (G) . Percentages and numbers of CD19+ B cells from the infected mice at 8 d.p.i. by flow cytometry and counting (n=4) (H) . Measurements of H1N1-specific IgG (I) and IgA (J) in the BALF of the CXCL5 treatment and control mice at 8 d.p.i. (n = 4). The error bars represent the SDs of 4 samples. * P < 0.05 based on Student’s t -test.

Article Snippet: The membranes were stained with a rabbit anti-mouse CXCR2 polyclonal antibody (ab65968, Abcam, UK) and a goat anti-rabbit IgG H&L (HRP) polyclonal antibody (ab6721, Abcam, UK).

Techniques: Infection, Virus, Control, Enzyme-linked Immunosorbent Assay, Expressing, Flow Cytometry, Recombinant