recombinant human cxcl1 gro alpha protein  (R&D Systems)

Journal: Cancer Science

Article Title: Tumor‐Associated Macrophage‐Derived CXCL1 Promotes Endometrial Cancer Progression Through the CXCR2/NF‐κB Pathway

doi: 10.1111/cas.70324

Figure Lengend Snippet: Recombinant CXCL1 (rCXCL1) promotes the proliferation, migration, and EMT of endometrial cancer (EC) cells (Ishikawa and HEC‐1B). (A) Membranes from a human cytokine antibody array comparing the supernatant of Ishikawa cells alone (control) and co‐cultured with M2 macrophages. The box highlights CXCL1 spots. (B) Representative western blotting images of EMT markers in EC cells treated with different concentrations of rCXCL1. (C–E) Proliferative capability of EC cells treated with rCXCL1 (20 and 40 ng/mL) assessed by CCK8 assay, colony formation assay, and Edu assay. (F) Migration capability of EC cells assessed by Transwell and wound‐healing assay. Data are presented as the mean ± SD. Statistical analysis was performed using Student's t ‐test. (** p < 0.01, *** p < 0.001, and **** p < 0.0001).

Article Snippet: Subsequently, recombinant human CXCL1 (rCXCL1; MedChemExpress, USA), CXCL1 neutralizing antibody (CXCL1‐Nab; R&D Systems, USA), or the NF‐κB inhibitor BAY 11‐7082 (MedChemExpress, USA) was introduced into the lower chamber.

Techniques: Recombinant, Migration, Ab Array, Control, Cell Culture, Western Blot, CCK-8 Assay, Colony Assay, EdU Assay, Wound Healing Assay

Journal: Cancer Science

Article Title: Tumor‐Associated Macrophage‐Derived CXCL1 Promotes Endometrial Cancer Progression Through the CXCR2/NF‐κB Pathway

doi: 10.1111/cas.70324

Figure Lengend Snippet: M2 macrophages secrete CXCL1 to promote the proliferation, migration and EMT of endometrial cancer (EC) cells (Ishikawa and HEC‐1B). (A, B) In the presence or absence of CXCL1 neutralizing antibody, proliferative capability of EC cells with co‐culture treatment or rCXCL1 assessed by CCK8 assay, colony formation assay. (C) Western blot analysis demonstrating the expression of EMT markers in EC cells. (D) Representative immunofluorescence images of CXCR2 in EC cells. (E) Proliferative capability of EC cells by Edu assay. (F) Migration capability of EC cells by Transwell assay and wound‐healing assay. Data are presented as the mean ± SD. Statistical analysis was performed using one‐way ANOVA. (**** p < 0.0001, *** p < 0.001 vs. control group. #### p < 0.001, ## p < 0.01 vs. co‐culture group).

Article Snippet: Subsequently, recombinant human CXCL1 (rCXCL1; MedChemExpress, USA), CXCL1 neutralizing antibody (CXCL1‐Nab; R&D Systems, USA), or the NF‐κB inhibitor BAY 11‐7082 (MedChemExpress, USA) was introduced into the lower chamber.

Techniques: Migration, Co-Culture Assay, CCK-8 Assay, Colony Assay, Western Blot, Expressing, Immunofluorescence, EdU Assay, Transwell Assay, Wound Healing Assay, Control

Journal: Cancer Science

Article Title: Tumor‐Associated Macrophage‐Derived CXCL1 Promotes Endometrial Cancer Progression Through the CXCR2/NF‐κB Pathway

doi: 10.1111/cas.70324

Figure Lengend Snippet: CXCL1 binds CXCR2 to promote the proliferation, migration, and EMT of endometrial cancer (EC) cells (Ishikawa and HEC‐1B). (A) The protein and mRNA levels of CXCR2 in EC cells with and without co‐culture treatment. (B) Western blot analysis demonstrating the expression of EMT markers in EC cells transfected with shCXCR2 under co‐culture or rCXCL1 treatment. (C–E) Proliferative capability of EC cells transfected with shCXCR2 under co‐culture or rCXCL1 treatment assessed by CCK8 assay, colony formation assay, and Edu assay. (F) Migration capability of EC cells treated as described above assessed by Transwell assay and wound‐healing assay. Data are presented as the mean ± SD. Statistical analysis was performed using one‐way ANOVA. (**** p < 0.0001, *** p < 0.001 vs. control group. ### p < 0.001, ## p < 0.01 vs. co‐culture group).

Article Snippet: Subsequently, recombinant human CXCL1 (rCXCL1; MedChemExpress, USA), CXCL1 neutralizing antibody (CXCL1‐Nab; R&D Systems, USA), or the NF‐κB inhibitor BAY 11‐7082 (MedChemExpress, USA) was introduced into the lower chamber.

Techniques: Migration, Co-Culture Assay, Western Blot, Expressing, Transfection, CCK-8 Assay, Colony Assay, EdU Assay, Transwell Assay, Wound Healing Assay, Control

Journal: Cancer Science

Article Title: Tumor‐Associated Macrophage‐Derived CXCL1 Promotes Endometrial Cancer Progression Through the CXCR2/NF‐κB Pathway

doi: 10.1111/cas.70324

Figure Lengend Snippet: CXCL1 and CXCR2 expression in normal endometrium and endometrial cancer (EC) tissues. (A) Western blot analysis the protein levels of CXCR2 in fresh EC tissues and adjacent non‐tumor tissues( n = 6). (B) Immunohistochemical staining of CXCL1 and CXCR2 in normal endometrium and EC (G1–G3). Scale bars represent 100 μm. (C) The proportion of positive areas in different groups. (D) The correlation between CD163+ cell numbers and positive areas in tumor tissue (Spearman's correlation). (E) Immunofluorescence localization of CXCR2 (red), CXCL1 (green), CD163 (red) and E‐cadherin (green) in EC. For primary antibodies from the same species, we used the Flexible Coralite 488 Antibody Labeling Kit according to the manufacturer's instructions. Data are presented as the mean ± SD. (*** p < 0.001).

Article Snippet: Subsequently, recombinant human CXCL1 (rCXCL1; MedChemExpress, USA), CXCL1 neutralizing antibody (CXCL1‐Nab; R&D Systems, USA), or the NF‐κB inhibitor BAY 11‐7082 (MedChemExpress, USA) was introduced into the lower chamber.

Techniques: Expressing, Western Blot, Immunohistochemical staining, Staining, Immunofluorescence, Antibody Labeling

Journal: Cancer Science

Article Title: Tumor‐Associated Macrophage‐Derived CXCL1 Promotes Endometrial Cancer Progression Through the CXCR2/NF‐κB Pathway

doi: 10.1111/cas.70324

Figure Lengend Snippet: M2 macrophages promote the EMT of endometrial cancer (EC) cells (Ishikawa and HEC‐1B) via the CXCR2/NF‐κB signaling pathway. (A) Western blot analysis demonstrating the expression of CXCR2 and NF‐κB pathway in EC cells under co‐culture or rCXCL1 treatment, with or without CXCL1 neutralizing antibody. (B) Translocation of NF‐κB from the cytoplasm to the nucleus was detected using immunofluorescence to indicate the activation of NF‐κB signaling in EC cells transfected with or without shCXCR2 under conditions of either co‐culture or rCXCL1 treatment. (C) Western blot analysis demonstrating the expression of CXCR2 and NF‐κB pathway in EC cells transfected with shCXCR2 under co‐culture or rCXCL1 treatment. (D) Western blot analysis showed that the NF‐κB inhibitor Bay 11–7082 inhibited the activation of the EMT process induced by co‐culture or rCXCL1.

Article Snippet: Subsequently, recombinant human CXCL1 (rCXCL1; MedChemExpress, USA), CXCL1 neutralizing antibody (CXCL1‐Nab; R&D Systems, USA), or the NF‐κB inhibitor BAY 11‐7082 (MedChemExpress, USA) was introduced into the lower chamber.

Techniques: Western Blot, Expressing, Co-Culture Assay, Translocation Assay, Immunofluorescence, Activation Assay, Transfection

Journal: Cancer Science

Article Title: Tumor‐Associated Macrophage‐Derived CXCL1 Promotes Endometrial Cancer Progression Through the CXCR2/NF‐κB Pathway

doi: 10.1111/cas.70324

Figure Lengend Snippet: Schematic illustration of the functional role of M2 macrophages and their secretion of CXCL1 in endometrial cancer (created by BioRender.com ).

Article Snippet: Subsequently, recombinant human CXCL1 (rCXCL1; MedChemExpress, USA), CXCL1 neutralizing antibody (CXCL1‐Nab; R&D Systems, USA), or the NF‐κB inhibitor BAY 11‐7082 (MedChemExpress, USA) was introduced into the lower chamber.

Techniques: Functional Assay

Journal: Cancer Science

Article Title: Tumor‐Associated Macrophage‐Derived CXCL1 Promotes Endometrial Cancer Progression Through the CXCR2/NF‐κB Pathway

doi: 10.1111/cas.70324

Figure Lengend Snippet: Neutralization of CXCL1 and inhibiting the NF‐κB pathway inhibited TAM‐induced progression of endometrial cancer in vivo. (A) A schematic diagram illustrating the animal experimentation protocol (created by BioRender.com ) with n = 5 mice per group. (B) The xenograft tumors in nude mice were removed and photographed. (C) The volume of the xenograft tumor was calculated, and a tumor growth curve was plotted. (D) Weight of xenografts tumors was measured and compared among four groups. (E) The protein level of E‐cadherin, Vimentin, CXCR2, and p‐P65 in xenograft tumors from four groups by western blot. (F) HE staining and immunohistochemical analysis of E‐cadherin, Vimentin, CXCR2, and p‐P65 in xenograft tumors from four groups. Data are presented as the mean ± SD. (* p < 0.05, ** p < 0.01 and *** p < 0.001).

Article Snippet: Subsequently, recombinant human CXCL1 (rCXCL1; MedChemExpress, USA), CXCL1 neutralizing antibody (CXCL1‐Nab; R&D Systems, USA), or the NF‐κB inhibitor BAY 11‐7082 (MedChemExpress, USA) was introduced into the lower chamber.

Techniques: Neutralization, In Vivo, Western Blot, Staining, Immunohistochemical staining