anti cxcl2 antibody  (R&D Systems)

Journal: Communications Biology

Article Title: The crosstalk of monocyte-neutrophil in hair follicles regulates neutrophil transepidermal migration in contact dermatitis

doi: 10.1038/s42003-025-07960-w

Figure Lengend Snippet: A ELISA detection of skin TNF-α production in PBS-Lipo or Clod-Lipo-treated group. n = 3, means ± SEM. B ELISA detection of skin CXCL2 production in PBS-Lipo or Clod-Lipo-treated group. n = 3, means ± SEM. C qPCR examination of Tnf expression in skin infiltrated cells. n = 9, means ± SEM. D qPCR examination of Cxcl2 expression in skin infiltrated cells. n = 9, means ± SEM. E Flow cytometry analysis of NTEM ratio in Tnf -deficient and Tnf -competent mice. n = 9, means ± SEM, 3 mice were excluded due to the technical complications. F qPCR analysis of Cxcl2 expression in Tnf -deficient and Tnf -competent mice. G qPCR analysis of Mmp9 expression Tnf -deficient and Tnf -competent mice. n = 9, means ± SEM. H Epidermis/dermis NTEM ratio in CXCR2 inhibitor or vehicle-treated group. n = 9, means ± SEM. I Epidermal/dermal infiltrated neutrophil ratio (NTEM ratio) in anti-CXCL2 or IgG treated group. n = 12, means ± SEM. J , qPCR analysis of Mmp9 expression in anti-CXCL2 or IgG treated group, n = 6–9. Data are representative of two independent experiments in ( C , D , J ). All the mice were female for these experiments. Data are representative of three independent experiments in ( E–I ). Statistical analysis was performed using a two-tailed unpaired Student’s t -test in ( A , B , E–J ). Data in ( C , D ) were analyzed using One-way ANOVA followed by Bonferroni’s post hoc test. Bonferroni-corrected p values are indicated in ( C , D ).

Article Snippet: To study the function of CXCL2 in NTEM, female mice (weighing ~20 g) were intravenously injected with 30 μg of anti-mouse CXCL2 monoclonal antibody (Cat. No. MAB452-500, R&D Systems, USA) or an isotype control (Rat IgG2b, clone: 141945, Cat. No. MAB0061, R&D Systems, USA) 6 h before ICD induction.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Flow Cytometry, Two Tailed Test

Journal: Communications Biology

Article Title: The crosstalk of monocyte-neutrophil in hair follicles regulates neutrophil transepidermal migration in contact dermatitis

doi: 10.1038/s42003-025-07960-w

Figure Lengend Snippet: A Representative flow cytometry images of skin infiltrated neutrophils in WT (BM WT), WT (BM Tnf −/− ), and Tnf -−/− (BM WT) groups. 6 Tnf -deficient male mice were used as BM donors, all the recipient mice were female. B Statistical flow cytometry analysis of epidermal/dermal infiltrated neutrophil ratio (NTEM ratio) in WT (BM WT), WT (BM Tnf −/− ), Tnf −/− (BM WT) group. n = 9, means ± SEM. C Quantification of gelatin zymography of invitro cultured neutrophils activated by TNF-α and CXCL2. n = 6, 3 male mice and 3 female mice were used to obtain BM and purify neutrophils in vitro, means ± SEM. D Images displaying gelatin zymography of ( C ). E Images display gelatin zymography of in vitro cultured neutrophils with dermal purified monocytes. F Quantification of gelatin zymography of in vitro cultured neutrophils with dermal purified monocytes in ( E ). n = 6, means ± SEM. G Quantification of gelatin zymography of in vitro cultured neutrophils with Tnf -competent-monocytes or Tnf -deficient-monocytes. I Images display gelatin zymography in ( G ). H Quantification of gelatin zymography of in vitro cultured neutrophils with siNC-monocytes or si Cxcl2 -monocyte. n = 6, means ± SEM. J Images display gelatin zymography in ( H ). The monocyte in ( G , H ) was collected from bone marrow and stimulated with LPS before coculture with neutrophils. The gelatin zymography in ( D , E , I , J ) are representative images of two or three independent trials. ND, not detected, ns, no significant difference. Data in ( B ) are from three independent trials; data in ( C , F , G , H ) are from two independent trials. Female mice were used to obtain BM in ( E–J ). Statistical analysis was performed using a two-tailed unpaired Student’s t -test in ( F , G , H ). Data in ( B , C ) were analyzed using a one-way ANOVA followed by Bonferroni’s post hoc test. Bonferroni-corrected p values are indicated in ( B , C ).

Article Snippet: To study the function of CXCL2 in NTEM, female mice (weighing ~20 g) were intravenously injected with 30 μg of anti-mouse CXCL2 monoclonal antibody (Cat. No. MAB452-500, R&D Systems, USA) or an isotype control (Rat IgG2b, clone: 141945, Cat. No. MAB0061, R&D Systems, USA) 6 h before ICD induction.

Techniques: Flow Cytometry, Zymography, Cell Culture, In Vitro, Purification, Two Tailed Test

Journal: Cell discovery

Article Title: Targeting a chemo-induced adaptive signaling circuit confers therapeutic vulnerabilities in pancreatic cancer.

doi: 10.1038/s41421-024-00720-w

Figure Lengend Snippet: Fig. 3 14-3-3ζ overexpressing PDAC cells increases CXCL2/5 via Yap1 in response to stresses. a Cytokine array analysis of CM from Panc02.shCtrl vs Panc02.shζ cells in 0% FBS culture for 72 h. b Left: WB analysis of CXCL2, CXCL5, and GAPDH (sample processing controls) expression in Panc02.shCtrl and Panc02.shζ cells cultured in 10% FBS or 0% FBS medium for 48 h. Right: WB analysis of CXCL2, CXCL5, and GAPDH (sample processing controls) in 3D cultured PANC-1.shCtrl and PANC-1.shζ cells treated with Gem (20 nM) vs vehicle for 48 h. c WB analysis of CXCL2, CXCL5, 14-3-3ζ, and GAPDH (sample processing controls) in PANC-1.shCtrl and PANC-1.shζ, and 14-3-3ζ-overexpressing PANC-1.shζ cells treated with Gem (20 nM) vs vehicle for 48 h. d WB analyses of Yap1, CXCL2, CXCL5, and GAPDH (sample processing controls) expression in Panc02.shCtrl and Panc02.shYap1 cells in 0% FBS culture for 24 h. Representative data of two independent repeats. e ChIP-qPCR assays of Yap1 binding to CXCL2/5 promoter region in 3D-cultured PANC-1 cells with 24 h of Gem (20 nM) treatment (mean ± SD, t-test, n = 3 biological repeats). f WB analysis of CXCL2, CXCL5, and GAPDH (sample processing controls) in Panc02.shCtrl and Panc02.shNLK sublines cultured in 0.1% FBS for 24 h. Representative data of two independent repeats. g Schematics (left) and relative numbers of proliferating (middle) and apoptotic (right) cells from 3D-cultured PATC53 cells treated with CM from 3D-cultured hPSCs that were activated by adding CM from 8.5 nM Gem-treated 3D-cultured PATC53 cells plus CXCL2 (1 μg/mL) or CXCL5 (3 μg/mL) blocking antibodies for 48 h (mean ± SD, t-test, n = 3 biological repeats). h Schematics and relative cell number of 3D-cultured KPC mT3 cells treated with CM from 3D-cultured mPSCs added with vehicle, recombinant CXCL2 (0.5 ng/mL), or recombinant CXCL5 (0.1 µg/mL) proteins for 48 h (mean ± SD, t-test, n = 3 biological repeats). i Stress-induced 14-3-3ζ-Yap1-CXCL2/5 pathway in PDAC cells activates fibroblasts, which turns on the adaptive response that enables PDAC cells to survive under stress conditions.

Article Snippet: The blocking antibodies to mouse CXCL2 (MAB452), mouse CXCL5 (MAB433), human CXCL5 (MAB254), and control antibody (MAB0061) were purchased from R&D Systems and human CXCL2 (311001) was purchased from Biolegend (California, USA).

Techniques: Expressing, Cell Culture, ChIP-qPCR, Binding Assay, Blocking Assay, Recombinant

Journal: Heliyon

Article Title: hsa_circ_0007755 competitively adsorbs miR-27b-3p to mediate CXCL2 expression and recruit Th1 cells to promote hypertrophic scars development

doi: 10.1016/j.heliyon.2024.e39169

Figure Lengend Snippet: CXCL2 is the target gene of miR-27b-3p. A: starbase predicted the potential binding sites of CXCL2 and miR-27b-3p. B: Western blot tested CXCL2 in human normal skin tissue and HS tissue. C: Western blot tested CXCL2 in normal skin tissue and HS tissue of mice. D: Western blot tested CXCL2 in HFF-1 and hHSFs. E-F: Dual luciferase reporter assay and RIP analyzed the targeting relationship between CXCL2 and miR-27b-3p. G: Western blot tested the effect of overexpressed miR-27b-3p on CXCL2 expression in hHSFs. Data were expressed as mean ± SD (N = 3). ∗ P < 0.05.

Article Snippet: Primary antibodies used were: CXCL2 (MAB452, R&D Systems), Ki-67 (M7240, Dako), phospho-p65 (3033, Cell Signaling Technology), α-smooth muscle actin (α-SMA) (A5228, MilliporeSigma), collagen I (ab34710, Abcam), and GAPDH (ab8245, Abcam).

Techniques: Binding Assay, Western Blot, Luciferase, Reporter Assay, Expressing

Journal: Heliyon

Article Title: hsa_circ_0007755 competitively adsorbs miR-27b-3p to mediate CXCL2 expression and recruit Th1 cells to promote hypertrophic scars development

doi: 10.1016/j.heliyon.2024.e39169

Figure Lengend Snippet: The promotion effect of overexpressed hsa_circ_0007755 on proliferation, invasion and inflammation of hHSFs is mitigated by knockdown of CXCL2. pcDNA 3.1-hsa_circ_0007755 and si-CXCL2 were co-transfected into hHSFs. A: Western blot tested CXCL2. B: CCK-8 assayed hHSFs proliferation. C: Flow cytometry measured apoptosis rate. D: Transwell assayed invasion ability. E: Western blot analyzed Ki-67, α-SMA, collagen I, and p-p65. F: Flow cytometry detected TH1/TH2 cell changes. G: ELISA measured cytokines TNF-α, INF-γ, IL-4, and IL-10 in TH1/TH2 cells. Data were expressed as mean ± SD (N = 3). ∗ P < 0.05.

Article Snippet: Primary antibodies used were: CXCL2 (MAB452, R&D Systems), Ki-67 (M7240, Dako), phospho-p65 (3033, Cell Signaling Technology), α-smooth muscle actin (α-SMA) (A5228, MilliporeSigma), collagen I (ab34710, Abcam), and GAPDH (ab8245, Abcam).

Techniques: Knockdown, Transfection, Western Blot, CCK-8 Assay, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Heliyon

Article Title: hsa_circ_0007755 competitively adsorbs miR-27b-3p to mediate CXCL2 expression and recruit Th1 cells to promote hypertrophic scars development

doi: 10.1016/j.heliyon.2024.e39169

Figure Lengend Snippet: Low hsa_circ_0007755 inhibits HS and regulates TH1/TH2 balance in mice. A: Western blot tested hsa_circ_0007755. B: Representative images of HE staining of mouse scar tissue. C: Flow cytometry analyzed changes in TH1/TH2 ratio in mouse blood samples. D: ELISA measured serum levels of TNF-α, INF-γ, IL-4, and IL-10. E: Western blot tested CXCL2, Ki-67, α-SMA, collagen I, and p-p65 in mouse scar tissue. Data were expressed as mean ± SD (n = 6). ∗ P < 0.05.

Article Snippet: Primary antibodies used were: CXCL2 (MAB452, R&D Systems), Ki-67 (M7240, Dako), phospho-p65 (3033, Cell Signaling Technology), α-smooth muscle actin (α-SMA) (A5228, MilliporeSigma), collagen I (ab34710, Abcam), and GAPDH (ab8245, Abcam).

Techniques: Western Blot, Staining, Flow Cytometry, Enzyme-linked Immunosorbent Assay