Journal: British journal of cancer

Article Title: Selective inhibition of HDAC6 promotes bladder cancer radiosensitization and mitigates the radiation-induced CXCL1 signalling.

doi: 10.1038/s41416-023-02195-0

Figure Lengend Snippet: Fig. 5 Tubacin represses RT-induced oncogenic CXCL1 signalling. a CXCL1 mRNA expression in T24 cells with or without 5 μM tubacin (Tub) treatment followed by 5 Gy irradiation (RT) was assessed by qPCR. Panobinostat (PAN, 10 nM) was used for comparison. b CXCL1 protein levels in conditioned medium from T24 cells with or without 5 μM Tub treatment followed by 5 Gy irradiation were measured by ELISA at 24 h. c Migration and invasion assays of T24 cells with or without 5 μM Tub treatment followed by 5 Gy irradiation. Migrated and invaded cells on the lower Transwell membrane were stained and calculated at 16 and 20 h, respectively. Representative images are shown below individually. d Migration and invasion assays of T24 cells treated with conditioned medium (CM) from cells with or without 5 μM Tub treatment followed by 5 Gy irradiation. Migrated and invaded cells on the lower Transwell membrane were stained and calculated at 16 and 20 h, respectively. Representative images are shown below individually. e Migration and invasion assays of Tub- and RT + Tub-treated T24 cells in the presence of 10 μg/mL anti-CXCL1 or control IgG treatment. Panobinostat (PAN)- and RT + PAN-treated cells were used for comparison. Migrated and invaded cells on the lower Transwell membrane were stained and calculated at 16 and 20 h, respectively. Representative images are shown below individually. f Western blotting for H3K9ac, acetyl-α-tubulin and Snail in Tub- and RT + Tub-treated T24 cells with or without anti-CXCL1 treatment at 24 h. Panobinostat (PAN)- and RT + PAN-treated cells were used for comparison. Snail protein expression was quantified by ImageJ. Data are presented as the means ± SD from three independent biological replicates. Student’s t-test was used for significant differences. *P < 0.05; **P < 0.01; ***P < 0.001. g Immunohistochemical staining for CXCL1 in tumour tissues obtained from urothelial carcinoma patients. Representative images are shown, and brown precipitates indicate positive signals. Scale bar represents 50 μm. h Overall survival analysis of 40 urothelial carcinoma patients categorised by CXCL1 protein levels. Statistical differences were assessed by the log-rank (Mantel–Cox) test. i Categorisation of CXCL1 expression in patients with high (pT3-4) and low (pT1-2) T stages.

Article Snippet: The conditioned medium was collected at 24 h and the CXCL1 protein level was determined using a Quantikine ELISA kit (Human CXCL1/GROα, R&D Systems, Minneapolis, MN) according to the manufacturer’s instructions.

Techniques: Expressing, Irradiation, Comparison, Enzyme-linked Immunosorbent Assay, Migration, Membrane, Staining, Control, Western Blot, Immunohistochemical staining

Journal: Nature Communications

Article Title: Histone lactylation increases CXCL1 expression for neutrophil infiltration and immune escape in pancreatic cancer

doi: 10.1038/s41467-026-69311-5

Figure Lengend Snippet: A The flowchart illustrates that RNA sequencing was conducted using two PDAC cell lines with or without 2DG treatment. Subsequently, the intersection of detected genes within the chemokine family in both cell lines is identified, and a heatmap is generated to display the log 2 fold change (2DG versus vehicle) of these genes. B , C Relative mRNA levels of Cxcl1 in PDAC cell lines following glycolysis inhibition, either by using 2DG or silencing LDH, were analyzed by qRT-PCR ( n = 3 independent experiments). D , E Relative protein levels of CXCL1 in PDAC cell lines following glycolysis inhibition, either by using 2DG or silencing LDH, were determined using ELISA assay ( n = 3 independent experiments). F Representative IHC images of subcutaneous tumors ( n = 6 mice for 2DG treatment, n = 5 mice for shLDH/shNTC groups) stained by CXCL1 antibody (scale bars = 100 μm). G Relative serum CXCL1 levels in mice treated with or without 2DG were measured by ELISA ( n = 6 mice per group). H Relative serum CXCL1 levels in healthy donors and PDAC patients were measured by ELISA (22 healthy samples and 27 PDAC samples). I Schematic diagram showing the in vitro migration assay: human/mouse neutrophils were co-incubated with the culture medium supernatant from PDAC cells treated with 2DG or LDH-knockdown. J Neutrophils were co-cultured with CD8 + T cells in different proportions, and the proliferation of CD8 + T cells was detected with the CFSE assay ( n = 6 biologically independent samples). K The migratory activity of neutrophils co-incubated with the culture medium supernatant from PDAC cells treated with shLDH and recombinant CXCL1 was analyzed by counting the penetrated cell numbers ( n = 3 biologically independent samples). L The neutrophils were treated with SX-682 or Navarixin for 1.5 h in advance, and the migratory activity of neutrophils co-incubated with the culture medium supernatant from PDAC cell lines treated with shLDH was analyzed by counting the penetrated cell numbers ( n = 3 biologically independent samples). M Representative luminescence images of the orthotopic tumor in mice and statistical analysis of MFI ( n = 5 mice per group). N Tumor-infiltrating neutrophils isolated from the orthotopic tumors were analyzed using flow cytometry ( n = 5 mice per group). Data represent mean ± SEM. Statistical analysis was conducted using the two-tailed unpaired Student’s t test. Source data are provided as a Source Data file.

Article Snippet: The concentration of CXCL1 from healthy individuals and PDAC patients’ plasma was measured using a Human CXCL1 ELISA Kit (Proteintech, #KE00133) according to the manufacturer’s instructions.

Techniques: RNA Sequencing, Generated, Inhibition, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Staining, In Vitro, Migration, Incubation, Knockdown, Cell Culture, CFSE Assay, Activity Assay, Recombinant, Isolation, Flow Cytometry, Two Tailed Test

Journal: Nature Communications

Article Title: Histone lactylation increases CXCL1 expression for neutrophil infiltration and immune escape in pancreatic cancer

doi: 10.1038/s41467-026-69311-5

Figure Lengend Snippet: A Lactylation levels of pan-lysine and different lysine sites in histone H3 of 7 pairs of PDAC tissue and adjacent normal tissue were detected by western blot. B Representative IHC images of clinical PDAC and adjacent normal tissue stained by H3K18la antibody ( n = 79 patients). C The diagram illustrating the targets of 2DG, shRNA, and rotenone in the glycolysis process. D , E Relative levels of H3K18la and PanKla in PDAC cells treated with gradient doses of 2DG ( D ) or rotenone ( E ) for 24 h were detected by western blot. F Relative levels of H3K18la and PanKla in subcutaneous tumors treated with or without 2DG were detected by western blot. ( n = 4 biologically independent samples per group). G H3K18la levels of subcutaneous tumors treated with or without 2DG were detected by immunofluorescence staining. H The H3K18la levels in PDAC cells treated with 2DG or sodium lactate were detected by western blot. I The H3K18la level in PANC1 and PANC02 cells following LDH-knockdown and sodium lactate treatment by western blot. J Schematic diagram of the CUT&Tag assay for PanKla or H3K18la in BxPC3 cells with or without 2DG treatment. K TSS heatmaps presenting the binding density of PanKla and H3K18la with or without 2DG treatment. L Venn diagram indicating the overlap of differentially expressed genes in TCGA-PAAD between PDAC and adjacent normal tissue, differentially expressed genes in RNA-seq after 2DG treatment, and differentially bound genes of H3K18la and PanKla after 2DG treatment. M Genome browser track analysis shows the PanKla and H3K18la levels in the Cxcl1 promoter region with or without 2DG treatments. N The Cxcl1 promoters that are bound by H3K18la and PanKla in PDAC cells were quantified using ChIP-qPCR assays ( n = 3 independent experiments). O Relative mRNA levels of Cxcl1 in PDAC cells treated with 2DG or sodium lactate were detected by qRT-PCR ( n = 3 independent experiments). P Representative IHC images stained by CXCL1 or H3K18la antibody and correlation analysis of H-scores of CXCL1 and H3K18la in PDAC samples ( n = 21 patients). Scale bar = 100 μm. Data represent mean ± SEM. Statistical analysis was conducted using the two-tailed unpaired Student’s t test ( N–O ) and two-tailed Pearson’s correlation analysis ( P ). Unless otherwise indicated, all western blots had three independent experimental repetitions with consistent results, where Histone H3, H3K9la, H3K14la, H3K18la, and PanKla blots are from parallel-processed separate gels (size conflict) with samples from the same experiment. Source data are provided as a Source Data file.

Article Snippet: The concentration of CXCL1 from healthy individuals and PDAC patients’ plasma was measured using a Human CXCL1 ELISA Kit (Proteintech, #KE00133) according to the manufacturer’s instructions.

Techniques: Western Blot, Staining, shRNA, Immunofluorescence, Knockdown, Binding Assay, RNA Sequencing, ChIP-qPCR, Quantitative RT-PCR, Two Tailed Test

Journal: Nature Communications

Article Title: Histone lactylation increases CXCL1 expression for neutrophil infiltration and immune escape in pancreatic cancer

doi: 10.1038/s41467-026-69311-5

Figure Lengend Snippet: A Molecular docking simulated the binding affinities of PCAF for acetyl-CoA and lactyl-CoA. B Co-IP was performed to confirm the interaction between H3K18la and PCAF in PANC1 and KPC cells. PCAF and H3K18la were detected on the same gel. C, D Western blot analysis demonstrates the level of H3K18la in PDAC cells with or without the treatment of PCAF inhibitors (bromosporine or Embelin). Histone H3 and H3K18la blots are from parallel-processed separate gels (size conflict) with samples from the same experiment. E Western blot analysis demonstrates the level of H3K18la in PDAC cells following the knockdown of Pcaf . Histone H3, PCAF, β-actin and H3K18la blots are from parallel-processed separate gels with samples from the same experiment. F Western blots of in vitro histone acetylation or lactylation assay, the samples as indicated. Blots are from parallel-processed separate gels with samples from the same experiment. G , H Relative Cxcl1 RNA and protein levels with the vehicle or bromosporine treatment were analyzed by qRT-PCR and ELISA ( n = 3 independent experiments), respectively. I Relative Cxcl1 RNA levels after Pcaf knockdown were analyzed by qRT-PCR ( n = 3 independent experiments). J A schematic diagram showing the orthotopic tumor construction in C57BL/6 J mice ( n = 5 mice per group in one experiment, 2 × 10 6 KPC-luc cells per mouse) with or without bromosporine treatment. K , L Image and weights of the orthotopic tumors in the experimental groups from ( J ) at the end of the experiments ( n = 5 mice per group). M Lactylation levels of H3K18 and PanK in orthotopic tumors with or without bromosporine treatment were detected by western blot ( n = 4 biologically independent samples per group). Histone H3, H3K18la, and PanKla blots are from parallel-processed separate gels (size conflict) with samples from the same experiment. N Serum CXCL1 levels in mice with or without bromosporine treatment were measured by ELISA ( n = 5 mice per group). O – R Tumor-infiltrating neutrophils, CD8 + T cells, GZMB + CD8 + T cells, and PD-1 + CD8 + T cells isolated from the orthotopic tumors were analyzed using flow cytometry. Representative graphs of flow cytometry (left panel) and statistical analysis of the cell ratio (right panel) ( n = 5 mice per group). S The migratory abilities of neutrophils co-incubated with the culture medium supernatant from shNTC/ Pcaf PDAC cells treated with recombinant CXCL1 were analyzed by counting the penetrated cell numbers ( n = 3 biologically independent samples). T Representative IHC images stained by CXCL1 or H3K18la antibody (scale bar = 100 μm, left panel), correlation analysis of H-scores of CXCL1 and H3K18la in PDAC samples ( n = 21 patients, right panel). Data represent mean ± SEM. Statistical analysis was conducted using the two-tailed unpaired Student’s t test ( G – I , L , and N – S ) and two-tailed Pearson’s correlation analysis ( T ). Unless otherwise indicated, all western blots had three independent experimental repetitions with consistent results. Source data are provided as a Source Data file.

Article Snippet: The concentration of CXCL1 from healthy individuals and PDAC patients’ plasma was measured using a Human CXCL1 ELISA Kit (Proteintech, #KE00133) according to the manufacturer’s instructions.

Techniques: Binding Assay, Co-Immunoprecipitation Assay, Western Blot, Knockdown, In Vitro, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Isolation, Flow Cytometry, Incubation, Recombinant, Staining, Two Tailed Test

Journal: Nature Communications

Article Title: Histone lactylation increases CXCL1 expression for neutrophil infiltration and immune escape in pancreatic cancer

doi: 10.1038/s41467-026-69311-5

Figure Lengend Snippet: A A schematic diagram showing the subcutaneous tumor model ( n = 6 mice per group in one experiment, 6 × 10 6 PANC02 cells per mouse) treated with bromosporine and anti-PD-1 antibody. B. The tumor volume growth curves of subcutaneous tumors from ( A ). C , D Image and weights of the subcutaneous tumors at the end point of experiments ( n = 6 mice per group). E The statistical analysis of the cell ratio of tumor-infiltrating neutrophils and CD8 + T cells isolated from subcutaneous tumors ( n = 3 mice per group). F Western blot analysis demonstrates H3K18la levels in the subcutaneous tumors ( n = 3 biologically independent samples per group) from ( A ). Histone H3 and H3K18la blots are from parallel-processed separate gels (size conflict) with samples from the same experiment. G Relative serum CXCL1 levels in mice treated with or without bromosporine/anti-PD-1 antibody were measured by ELISA ( n = 6 mice per group). H A schematic diagram showing the combinational treatment schedule for the orthotopic KPC-luc tumor model ( n = 21 mice per group in one experiment, 2 × 10 6 KPC-luc cells per mouse). I , J Image and weights of the orthotopic tumors at the end point of experiments ( n = 6 mice per group). K – N Tumor-infiltrating neutrophils, CD8 + T cells, GZMB + CD8 + T cells, and PD-1 + CD8 + T cells were analyzed using flow cytometry. Representative graphs of flow cytometry (left panel) and statistical analysis of the cell ratio (right panel) ( n = 6 mice per group). O Western blot analysis showing the H3K18la levels in the orthotopic tumors ( n = 3 biologically independent samples per group) from ( H ). Histone H3 and H3K18la blots are from parallel-processed separate gels (size conflict) with samples from the same experiment. P Representative luminescence images and Quantification of radiance intensity of the mouse model in ( H ). Q Survival probability of mice with orthotopically transplanted PDAC ( n = 15 mice per group). R A working model displaying the signaling pathway through which the aerobic glycolysis-mediated Lactate-PCAF-H3K18la-CXCL1 axis modulates the tumor microenvironment in pancreatic cancer, and the scientific basis for the development of a novel therapeutic strategy for PDAC. Data represent mean ± SEM. Statistical analysis was conducted using the two-tailed unpaired Student’s t test ( B , D , E , G , J – N , P ) and the log-rank test ( Q ). Source data are provided as a Source Data file.

Article Snippet: The concentration of CXCL1 from healthy individuals and PDAC patients’ plasma was measured using a Human CXCL1 ELISA Kit (Proteintech, #KE00133) according to the manufacturer’s instructions.

Techniques: Isolation, Western Blot, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Two Tailed Test

Journal: Oncology Letters

Article Title: Enhanced expression of CXCL1 in renal cell carcinoma facilitates tumor cell malignancy via PI3K/AKT-dependent mechanisms

doi: 10.3892/ol.2025.15372

Figure Lengend Snippet: Increased expression of CXCL1 mRNA levels is associated with the clinical characteristics of patients with RCC. (A) The expression profiles of CXCL1 mRNA in normal tissues compared with ccRCC or pRCC tissues. The different levels of expression of CXCL1 mRNA within (B) different clinical grades and (C) subtypes of ccRCC tissues. Expression levels of CXCL1 mRNA in the (D) GSE53757 and the (E) GSE40435 datasets. The different levels of expression of CXCL1 mRNA within different (F) clinical grades and (G) stages of ccRCC tissues. Analysis of the expression level of CXCL1 within the clinical (H) T-status, (I) N-status and (J) M-status categories in ccRCC tissues. (K-M) A prognostic assessment was performed among the various groups. **P<0.01 vs. normal; # P<0.01 vs. normal, grade 1 and 2; ▲ P<0.01 vs. normal and ccA subtype. CXCL1, CXC motif chemokine ligand 1; RCC, renal cell carcinoma; ccRCC, clear cell RCC; pRCC, papillary RCC; T, tumor; N, lymph node; M, metastasis; OS, overall survival; DSS, disease-specific survival; PFS, progression-free survival.

Article Snippet: Following 24 h culture, the supernatant was isolated via centrifugation at 4°C, 1,000 × g for 5 min, and a Human CXCL1 ELISA kit (cat. no. EK0722; Wuhan Boster Biological Technology, Ltd.) was employed to determine and quantify the level of CXCL1 in the supernatant, following precisely the manufacturer's instructions.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Enhanced expression of CXCL1 in renal cell carcinoma facilitates tumor cell malignancy via PI3K/AKT-dependent mechanisms

doi: 10.3892/ol.2025.15372

Figure Lengend Snippet: Expression levels of CXCL1 mRNA are correlated with the expression of genes implicated in tumorigenesis, recruitment of immune cells and presence of MSI in renal cell carcinoma. (A) Identification of 10 hub genes exhibiting co-expression with CXCL1. (B) Analysis of the protein-interaction network involving CXCL1. (C) Analysis of the correlation between CXCL1 and the aforementioned 10 hub genes. (D) Analysis of the correlation between CXCL1 expression and immune cell recruitment. The correlation between CXCL1 expression and the (E) number of mutations is presented, as well as (F) MSI scores. (G) The expression levels of CXCL1 in the MSI group compared with the MSS group. *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001. CXCL, CXC motif chemokine ligand; ACKR1, atypical chemokine receptor 1; CCR, C-C chemokine receptor type 2; MSI, microsatellite instability; MSS, microsatellite stable or no apparent MSI; TIMER, tumor immune estimation resource; TCGA, The Cancer Genome Atlas; T, tumor; N, normal.

Article Snippet: Following 24 h culture, the supernatant was isolated via centrifugation at 4°C, 1,000 × g for 5 min, and a Human CXCL1 ELISA kit (cat. no. EK0722; Wuhan Boster Biological Technology, Ltd.) was employed to determine and quantify the level of CXCL1 in the supernatant, following precisely the manufacturer's instructions.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Enhanced expression of CXCL1 in renal cell carcinoma facilitates tumor cell malignancy via PI3K/AKT-dependent mechanisms

doi: 10.3892/ol.2025.15372

Figure Lengend Snippet: Protein levels of CXCL1 expression are markedly elevated in the tissues of patients with RCC. Representative images, illustrating strong-intensity staining of CXCL1 in (A) grade I, (B) grade II and (C) grade III RCC tissues. (D) Representative image, illustrating weak-intensity staining of CXCL1 in paracancerous tissues. CXCL1, CXC motif chemokine ligand 1; RCC, renal cell carcinoma.

Article Snippet: Following 24 h culture, the supernatant was isolated via centrifugation at 4°C, 1,000 × g for 5 min, and a Human CXCL1 ELISA kit (cat. no. EK0722; Wuhan Boster Biological Technology, Ltd.) was employed to determine and quantify the level of CXCL1 in the supernatant, following precisely the manufacturer's instructions.

Techniques: Expressing, Staining

Journal: Oncology Letters

Article Title: Enhanced expression of CXCL1 in renal cell carcinoma facilitates tumor cell malignancy via PI3K/AKT-dependent mechanisms

doi: 10.3892/ol.2025.15372

Figure Lengend Snippet: Exogenous CXCL1 treatment enhances the malignant phenotype of renal cell carcinoma cells. Assessment of cell proliferation using a Cell Counting Kit-8 assay in (A) 786-O and (B) CAKI-2 cells. Representative images from Transwell assays with (C) 786-O and (D) CAKI-2 cells (magnification, ×100). Assessment of cell migration using a Transwell assay in (E) 786-O and (F) CAKI-2 cells. *P<0.05; **P<0.01 vs. 0 ng/ml. CXCL1, CXC motif chemokine ligand 1.

Article Snippet: Following 24 h culture, the supernatant was isolated via centrifugation at 4°C, 1,000 × g for 5 min, and a Human CXCL1 ELISA kit (cat. no. EK0722; Wuhan Boster Biological Technology, Ltd.) was employed to determine and quantify the level of CXCL1 in the supernatant, following precisely the manufacturer's instructions.

Techniques: Cell Counting, Migration, Transwell Assay

Journal: Oncology Letters

Article Title: Enhanced expression of CXCL1 in renal cell carcinoma facilitates tumor cell malignancy via PI3K/AKT-dependent mechanisms

doi: 10.3892/ol.2025.15372

Figure Lengend Snippet: Overexpression of CXCL1 stimulates the malignant phenotype of renal cell carcinoma cells. Evaluation of CXCL3 expression in the supernatant from cell medium using ELISA assay in (A) 786-O and (B) CAKI-2 cells. Assessment of cell proliferation using a Cell Counting Kit-8 assay with (C) 786-O and (D) CAKI-2 cells. Representative images from Transwell assay experiments with (E) 786-O and (F) CAKI-2 cells (magnification, ×100). Assessment of cell migration using Transwell assay with (G) 786-O and (H) CAKI-2 cells. **P<0.01 vs. control. CXCL1, CXC motif chemokine ligand 1.

Article Snippet: Following 24 h culture, the supernatant was isolated via centrifugation at 4°C, 1,000 × g for 5 min, and a Human CXCL1 ELISA kit (cat. no. EK0722; Wuhan Boster Biological Technology, Ltd.) was employed to determine and quantify the level of CXCL1 in the supernatant, following precisely the manufacturer's instructions.

Techniques: Over Expression, Expressing, Enzyme-linked Immunosorbent Assay, Cell Counting, Transwell Assay, Migration, Control

Journal: Oncology Letters

Article Title: Enhanced expression of CXCL1 in renal cell carcinoma facilitates tumor cell malignancy via PI3K/AKT-dependent mechanisms

doi: 10.3892/ol.2025.15372

Figure Lengend Snippet: Low expression of CXCL1 suppresses the malignant phenotype of renal cell carcinoma cells. Evaluation of CXCL3 expression in the supernatant from cell medium using ELISA assay with (A) 786-O and (B) CAKI-2 cells. Assessment of cell proliferation using a Cell Counting Kit-8 assay with (C) 786-O and (D) CAKI-2 cells. Representative images from Transwell assay experiments (magnification, ×100) with (E) 786-O and (F) CAKI-2 cells. Assessment of cell migration using Transwell assay with (G) 786-O and (H) CAKI-2 cells. **P<0.1 01 vs. control. CXCL1, CXC motif chemokine ligand 1; siRNA, small interfering RNA.

Article Snippet: Following 24 h culture, the supernatant was isolated via centrifugation at 4°C, 1,000 × g for 5 min, and a Human CXCL1 ELISA kit (cat. no. EK0722; Wuhan Boster Biological Technology, Ltd.) was employed to determine and quantify the level of CXCL1 in the supernatant, following precisely the manufacturer's instructions.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Cell Counting, Transwell Assay, Migration, Control, Small Interfering RNA

Journal: Oncology Letters

Article Title: Enhanced expression of CXCL1 in renal cell carcinoma facilitates tumor cell malignancy via PI3K/AKT-dependent mechanisms

doi: 10.3892/ol.2025.15372

Figure Lengend Snippet: Overexpression of CXCL1 modulates the expression of PI3K/AKT pathway-associated proteins in renal cell carcinoma cells. Representative images from western blotting assays in (A) 786-O and (B) CAKI-2 cells. Semi-quantification of the protein expression levels of Bax, Bcl-2, PI3K and AKT in (C) 786-O and (D) CAKI-2 cells. **P<0.01 vs. control. CXCL1, CXC motif chemokine ligand 1; Bax, Bcl-2-associated X protein; Bcl-2, B-cell lymphoma-2; PI3K, phosphoinositide 3-kinase.

Article Snippet: Following 24 h culture, the supernatant was isolated via centrifugation at 4°C, 1,000 × g for 5 min, and a Human CXCL1 ELISA kit (cat. no. EK0722; Wuhan Boster Biological Technology, Ltd.) was employed to determine and quantify the level of CXCL1 in the supernatant, following precisely the manufacturer's instructions.

Techniques: Over Expression, Expressing, Western Blot, Control

Journal: Oncology Letters

Article Title: Enhanced expression of CXCL1 in renal cell carcinoma facilitates tumor cell malignancy via PI3K/AKT-dependent mechanisms

doi: 10.3892/ol.2025.15372

Figure Lengend Snippet: Blocking AKT reverses the promoting effect of overexpression of CXCL1 on the malignant behaviors of renal cell carcinoma cells. Assessment of cell proliferation using a Cell Counting Kit-8 assay in (A) 786-O and (B) CAKI-2 cells. (Representative images from Transwell assay experiments (magnification, ×100) with (C) 786-O and (D) CAKI-2 cells. Assessment of cell migration using Transwell assay with (E) 786-O and (F) CAKI-2 cells. (magnification, ×100). **P<0.01 compared with overexpression. CXCL1, CXC motif chemokine ligand 1.

Article Snippet: Following 24 h culture, the supernatant was isolated via centrifugation at 4°C, 1,000 × g for 5 min, and a Human CXCL1 ELISA kit (cat. no. EK0722; Wuhan Boster Biological Technology, Ltd.) was employed to determine and quantify the level of CXCL1 in the supernatant, following precisely the manufacturer's instructions.

Techniques: Blocking Assay, Over Expression, Cell Counting, Transwell Assay, Migration