cxcl5  (Bioss)

Journal: Nature Communications

Article Title: Residual tumor cells after insufficient radiofrequency ablation promote lung metastasis by educating CD177 hi PAD4 hi neutrophils

doi: 10.1038/s41467-025-66897-0

Figure Lengend Snippet: A Differences in RNA-seq between Ctrl and iRFA on day 3 post-iRFA ( n = 3 mice). Volcano plot of DEGs identified by DESeq2, based on a negative binomial model. B Cxcl5 ( P < 0.001) and Il1b ( P = 0.045) in CT26 cells ± OA treated at 37 °C/45 °C via ELISA ( n = 3 biological replicates). ANOVA. C Cxcl5 in CT26 cells treated with DMSO, SB190, or SB580 at 37 °C/45 °C ± OA ( n = 3 biological replicates). P < 0.001, ANOVA. #, P < 0.001 vs . 45 °C + OA + DMSO group. D MPO-dsDNA levels in neutrophils co-cultured with conditioned media from CT26 cells (37 °C/45 °C) ± OA, with Cxcl5 and / or CXCR1/2 inhibitor (Reparixin) treatment ( n = 3 biological replicates). P < 0.001, ANOVA. # vs . 37 °C; ▼ vs . 37 °C + OA; ★ vs . 45 °C; P < 0.001. E (Left) Immunofluorescence in neutrophils co-cultured with 37 °C and 45 °C + OA CT26 / DLD1 TCM supplemented with DMSO / Trametinib (MEK/ERK inhibitor). (Right) cit-H3 + area quantification ( n = 3 biological replicates). P < 0.001, ANOVA. F , Erk and pErk in the neutrophils from iRFA mice. G–I Proteins in neutrophils treated with 37 °C or 45 °C ± OA CT26 TCM supplemented with ± ( G ) Cxcl5, ( H ) SB190, and ( I ) Reparixin. J Lung tissues from the Ctrl and iRFA mice treated with DMSO, Trametinib and/or a P38 inhibitor (Ralimetinib). K HE staining of lung tissues (Circles: lung metastases). L The quantification of lung metastatic area ( n = 9 mice). Ctrl groups: P = 0.059; iRFA groups: P < 0.001; ANOVA. M (Left) Ly6G and cit-H3 immunofluorescence in liver tumors. (Right) Quantification of the cit-H3 + area ( n = 9 mice). P < 0.001, ANOVA. Scale bars, 200 μm. N Ly6G and pErk immunofluorescence in liver tumors. Scale bars, 50 μm. Data are presented as mean values ± SD. C , H SB190: SB202190. C SB580: SB203580. E 37/45: 37 °C/45 °C. E and G – I TCM: conditioned media from tumor cells. Source data are provided as a Source Data file.

Article Snippet: After dewaxing, hydration, and antigen retrieval, the slices were incubated with the following primary antibodies: mCherry (1:200, ab167453, Abcam), Cd45 (1:200, GB11066, Servicebio), CD36 (1:200, 18836-1-AP, Proteintech), FABP4 (1:200, 12802-1-AP, Proteintech), FABP5 (1:200, 12348-1-AP, Proteintech), PPARγ (1:200, 10156-2-AP, Proteintech), p-P38 (1:200, 4511 T, Cell Signaling Technology), and CXCL5 (1:250, bs-2549R, Bioss).

Techniques: RNA Sequencing, Enzyme-linked Immunosorbent Assay, Cell Culture, Immunofluorescence, Staining

Journal: Nature Communications

Article Title: Residual tumor cells after insufficient radiofrequency ablation promote lung metastasis by educating CD177 hi PAD4 hi neutrophils

doi: 10.1038/s41467-025-66897-0

Figure Lengend Snippet: A Treatment timeline for a representative CRLM patient. The patient underwent Dixon surgery following CRLM diagnosis. First tumor evaluation showed a new metastasis in liver S5, and subsequently, RFA was performed. Postoperative PET/CT showed no tumor viability. Intrahepatic S7 metastasis appeared 4 months later. S5 and S7 tissues were obtained through hepatic resection, and pathological examination as well as sequencing were performed. The second progression with new lung metastases occurred at 7 months. Triangles: liver or lung metastases. B Neutrophil infiltration levels in tumors pre- and post-iRFA analyzed by different datasets (Box center line: median; box limits: upper and lower quartiles; whiskers: 1.5× interquartile range). C Representative GSEA analysis of FA metabolism and the PPAR pathway, and the differences in scRNA-seq data between pre- and post-iRFA tumors. Kolmogorov-Smirnov test. FDR correction. D CD36 and CXCL5 staining of the tumors pre- and post-iRFA. E Density quantification of CD36 + and CXCL5 + areas in the tumors pre- and post-iRFA. F CD66b immunofluorescence in tumors pre- and post-iRFA. Scale bars, 50 μm and 10 μm. CD66b + neutrophil counts shown on the left. G MPO and cit-H3 immunofluorescence in tumors pre- and post-iRFA. Scale bars, 50 μm. The cit-H3 + area shown on the left. H Mesenchymal markers, epithelial markers, and CD36 immunofluorescence on CTCs were analyzed. The data represented the proportions of CD36 hi -CTCs, CD36 hi -E-CTCs, and CD36 hi -EM-CTCs in the tumors pre- and post-iRFA. Scale bars, 10 μm. I MPO-dsDNA levels in the peripheral blood pre- and post-iRFA. J The data represented the proportions of CD36 hi -CTCs and CD36 hi -EM-CTCs in the cRFA and iRFA tumors. K MPO-dsDNA levels in the peripheral blood of cRFA and iRFA patients. Data are presented as mean values ± SD; Statistical analysis was performed based on Paired t -test ( B , E , F , and G , n = 8 patients; H and I , n = 13 patients) or Unpaired two-sided t -test ( J , K ); cRFA, n = 5 patients, iRFA, n = 13 patients). Source data are provided as a Source Data file.

Article Snippet: After dewaxing, hydration, and antigen retrieval, the slices were incubated with the following primary antibodies: mCherry (1:200, ab167453, Abcam), Cd45 (1:200, GB11066, Servicebio), CD36 (1:200, 18836-1-AP, Proteintech), FABP4 (1:200, 12802-1-AP, Proteintech), FABP5 (1:200, 12348-1-AP, Proteintech), PPARγ (1:200, 10156-2-AP, Proteintech), p-P38 (1:200, 4511 T, Cell Signaling Technology), and CXCL5 (1:250, bs-2549R, Bioss).

Techniques: Biomarker Discovery, Positron Emission Tomography-Computed Tomography, Sequencing, Staining, Immunofluorescence

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: a Cytokine array analysis of CM from Panc02.shCtrl vs Panc02.sh ζ 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.sh ζ 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.sh ζ 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.sh ζ 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.sh Yap1 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.sh NLK 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: Antibody to mouse CXCL5 (bs-2549R) was from Bioss antibodies (Massachusetts, USA).

Techniques: Expressing, Cell Culture, Binding Assay, Blocking Assay, Recombinant

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: a WB analyses of Cox2 and GAPDH (sample processing controls) in mPSCs (3D culture) incubated with recombinant CXCL2 (0.5 ng/mL) or CXCL5 (0.1 µg/mL) for 24 h. Representative data of two independent repeats. b WB analysis of Cox2 and GAPDH (sample processing controls) in NIH3T3 cells treated with CM from Panc02.shCtrl, Panc02.sh CXCL2 , or Panc02.sh CXCL5 cells cultured in 0% FBS for 72 h. Representative data of two independent repeats. c PGE2 concentration in CM from NIH3T3 cells treated with recombinant CXCL2 (0.5 ng/mL) or CXCL5 (0.1 µg/mL) for 48 h (mean ± SD, t -test, n = 3 biological repeats). d Relative numbers of Panc02 cells treated with PGE2 in 0% FBS for 48 h (mean ± SD, t -test, n = 3 biological repeats). e Relative numbers of PANC-1 cells in upper chambers of Transwell units 3D-cocultured with hPSCs in lower chambers of Transwell units with or without Cel (4 nM) and Gem (20 nM) for 72 h (mean ± SD, t -test, n = 3 biological repeats). f Relative number of Panc02 cells treated with CM from WT mPSCs or Cox2 –/– mPSCs activated by CM from Panc02 cells in 0% FBS for 48 h (mean ± SD, t -test, n = 3 biological repeats). g RPPA analysis of NIH3T3 cells treated with CM collected from Panc02.shCtrl and Panc02.sh ζ cells cultured in 10% or 0% FBS medium. h WB analysis of pT346-NDRG1, NDRG1, and GAPDH (sample processing controls) in 3D-cultured hPSCs treated for 24 h with CM collected from 3D-cultured PANC-1.shCtrl and PANC-1.sh ζ cells treated with vehicle/Gem (20 nM) for 24 h. Representative data of two independent repeats. i WB analysis of pT346-NDRG1, NDRG1, pS473-Akt, Akt, and GAPDH (sample processing controls) expression in 3D-cultured hPSCs treated with recombinant CXCL2 (15 ng/mL) or CXCL5 (25 ng/mL) for 24 h. Representative data of two independent repeats. j WB analysis of Rictor, pT346-NDRG1, NDRG1, Cox2, and GAPDH (sample processing controls) protein in NIH3T3.shCtrl and NIH3T3.sh Rictor cells treated for 24 h with recombinant CXCL2 (0.5 ng/mL) or recombinant CXCL5 (0.1 µg/mL). Representative data of two independent repeats. k, l IHC staining of indicated proteins of PDAC tumor tissues from KPC and KPC -ζ fl/fl mice with indicated treatments (mean ± SD, t -test, n = 5 biological repeats, scale bar: 20 µm). m PDAC cells and fibroblasts together create the adaptive stress response circuit, which is essential for PDAC cell adaptation to stresses, including nutrient deprivation and chemotherapy-induced genotoxicity. Stressors promote the Yap1-CXCL2/5 signaling axis via NLK in 14-3-3ζ-overexpressing PDAC cells, leading to the activation of the CXCR2-mTORC2-Cox2-PGE2 pathway in fibroblasts. Reciprocally, PGE2 from fibroblasts promotes PDAC cell survival and adaptive resistance to Gem.

Article Snippet: Antibody to mouse CXCL5 (bs-2549R) was from Bioss antibodies (Massachusetts, USA).

Techniques: Incubation, Recombinant, Cell Culture, Concentration Assay, Expressing, Immunohistochemistry, Activation Assay