primary antibodies against cxcl1  (R&D Systems)

Journal: Frontiers in Oncology

Article Title: Identification of immune-associated genes for the diagnosis of ulcerative colitis-associated carcinogenesis via integrated bioinformatics analysis

doi: 10.3389/fonc.2024.1475189

Figure Lengend Snippet: Verification and ROC curves of the hub genes. (A–D) The expression verification in the GSE87466 dataset. (E–H) Hub gene expression in the GSE39582 dataset. (I–L) ROC curves of BCL6, CXCL1, LCN2 and CXCL2 in the GSE92415 dataset. (M–P) ROC curves of BCL6, CXCL1, LCN2 and CXCL2 in the GSE20916 dataset. *p<0.05, ***p<0.001.

Article Snippet: The membrane was washed twice with TBST for 5 minutes each and subsequently incubated overnight at 4°C with primary antibodies against CXCL1 (1:500, 12335-1-AP, Proteintech), BCL6 (1:1000, 66340-1-Ig, Proteintech), and β-actin (1:1000, 7A-09, ab8226, ZSGB-BIO).

Techniques: Expressing, Gene Expression

Journal: Frontiers in Oncology

Article Title: Identification of immune-associated genes for the diagnosis of ulcerative colitis-associated carcinogenesis via integrated bioinformatics analysis

doi: 10.3389/fonc.2024.1475189

Figure Lengend Snippet: Verification of gene expression in colon tissues (A) Images of a normal intestinal tract obtained via white light endoscopy. (B) Intestinal tract images of UC patients via white light endoscopy. (C) Intestinal tract images of CRC patients via white light endoscopy. (D) NBI images of the intestinal tract of CRC patients. (E, H) H&E staining of human normal colon sections. (F, I) H&E staining of intestinal tissues from UC patients. (G, J) H&E staining of intestinal sections from CRC patients. (K–P) Expression of CXCL1 (K–M) and BCL6 (N–P) in the control cohort, UC patient cohort and cancer patient cohort determined by IHC analysis. (S, T) Statistical comparison of the IHC analysis of the mean densities of CXCL1 (S) and BCL6 (T) in the control cohort, UC patient cohort and cancer patient cohort. CXCL1 was expressed at the highest level in the cancer patient cohort. BCL6 was expressed at the highest level in the UC patient cohort. (Q, R) Western blot analysis of CXCL1 and BCL6 expression in human intestinal tissues. In the cancer patients group, the CXCL1 expressed the highest than others. In the UC patient group, the BCL6 expression was greater than that in the other groups. *p<0.05, **p<0.01, ***p<0.001 and ****p<0.0001.

Article Snippet: The membrane was washed twice with TBST for 5 minutes each and subsequently incubated overnight at 4°C with primary antibodies against CXCL1 (1:500, 12335-1-AP, Proteintech), BCL6 (1:1000, 66340-1-Ig, Proteintech), and β-actin (1:1000, 7A-09, ab8226, ZSGB-BIO).

Techniques: Gene Expression, Staining, Expressing, Control, Comparison, Western Blot

Journal: Frontiers in Oncology

Article Title: Identification of immune-associated genes for the diagnosis of ulcerative colitis-associated carcinogenesis via integrated bioinformatics analysis

doi: 10.3389/fonc.2024.1475189

Figure Lengend Snippet: Immunofluorescence images of CXCL1 and BCL6. (A–I) Immunofluorescence images of CXCL1 in colon tissues from the control, UC and cancer cohorts. (J) Quantitative analysis of CXCL1 immunofluorescence. (K–S) Immunofluorescence images of BCL6 in colon tissues from the control, UC and cancer groups. (T) Quantitative analysis of BCL6 immunofluorescence. ***p<0.001 and ****p<0.0001.

Article Snippet: The membrane was washed twice with TBST for 5 minutes each and subsequently incubated overnight at 4°C with primary antibodies against CXCL1 (1:500, 12335-1-AP, Proteintech), BCL6 (1:1000, 66340-1-Ig, Proteintech), and β-actin (1:1000, 7A-09, ab8226, ZSGB-BIO).

Techniques: Immunofluorescence, Control

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Breaking the symmetry of cell contractility drives tubulogenesis via CXCL1 polarization.

doi: 10.1073/pnas.2315894121

Figure Lengend Snippet: Fig. 3. Soft matrix induces CXCL1 polarization promoting cell protrusions and tubulogenesis. Analysis of (A) Cxcl1 mRNA expression of cells grown on the fish skin–derived collagen gel–coating or gel for 6 h and their (B) CXCL1 protein level in the culture supernatant. **P < 0.01; ***P < 0.001. (C) Representative confocal microscopy images showing CXCL1 (green), pT18S19MLC2 (red), microtubules (cyan), and nucleus (blue) in cells on the fish skin–derived collagen gel or gel-coating for 6 h. (D) Statistical analysis of mean fluorescence intensity of CXCL1 per high power field (HPF) in C. There were approximately 61 and 104 cells per HPF in the gel-coating and gel group, respectively. ***P < 0.001. Analysis of (E) Cxcl1 mRNA expression of cells grown on the 9-mo-old rat tail collagen gel with or without sonication (75% amplitude) for 1 d and their (F) CXCL1 protein level in the culture supernatant. ***P < 0.001. (G) Representative epifluorescent microscopy images of cellular staining for microtubules (green), microfilaments (red), and nucleus (blue) in cells treated with SB225002 on the fish skin–derived collagen gel for 7 h (yellow arrows: cell protrusions). (H) Montage images extracted from timelapse recordings of cells treated with siRNAs on fish skin–derived collagen gel (Movie S4) (red arrows; cell protrusions at 6 h). (I) Statistical analysis of relative cell protrusion activity per high power field (HPF) in Fig. 4H. There were approximately 310, 267, and 247 cells per HPF at 1 h in the siCntl, siCxcl1, and siCxcr1 group, respectively. **P < 0.01. (J) Tube formation on fish skin–derived collagen gel for 2 d in cells with siRNA knockdown of CXCL1. Cells with siCntl served as control. ***P < 0.001. (K) Tube formation on fish skin–derived collagen gel for 1 d in cells treated with SB225002 CXCRs inhibitor. *P < 0.05; **P < 0.01. (L) Tube formation on the fish skin–derived collagen gel for 2 d in cells with siRNA knockdown of CXCR1. ***P < 0.001.

Article Snippet: Immunofluorescent staining was performed using specific antibodies against CXCL1 (AF- 515- NA, R&D Systems), tubulin- α (05- 829, Merck), pMLC2 (95777, Cell Signaling Technology), and E- cadherin (610182, BD Biosciences) followed by Alexa Fluor- conjugated secondary antibodies (Invitrogen).

Techniques: Expressing, Derivative Assay, Confocal Microscopy, Fluorescence, Sonication, Microscopy, Staining, Activity Assay, Knockdown, Control

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Breaking the symmetry of cell contractility drives tubulogenesis via CXCL1 polarization.

doi: 10.1073/pnas.2315894121

Figure Lengend Snippet: Fig. 4. Reduction of cell contractility promotes cell protrusions. (A) Real-time PCR analysis of Cxcl1 expression in cells treated with or without blebbistatin on fish skin–derived collagen gel for 1 d. (B) ELISA of CXCL1 protein expression in the cell culture supernatant from cells treated with or without blebbistatin on fish skin–derived collagen gel for 1 d. (C) Representative confocal microscopy images showing CXCL1 (green), microtubules (red), microfilaments (cyan), and nucleus (blue) in cells treated with or without blebbistatin and grown on fish skin–derived collagen gel for 6 h. (D) Representative epifluorescent microscopy images of CXCL1 (green), microtubules (red), and nucleus (blue) in cells treated with or without blebbistatin on fish skin–derived collagen gel for 6 h. Cell morphology was observed through the bright field channel (Ph) (yellow arrows: cell protrusions). (E) Statistical analysis of cell protrusions per high-power field (HPF) of D. ***P < 0.001.

Article Snippet: Immunofluorescent staining was performed using specific antibodies against CXCL1 (AF- 515- NA, R&D Systems), tubulin- α (05- 829, Merck), pMLC2 (95777, Cell Signaling Technology), and E- cadherin (610182, BD Biosciences) followed by Alexa Fluor- conjugated secondary antibodies (Invitrogen).

Techniques: Real-time Polymerase Chain Reaction, Expressing, Derivative Assay, Enzyme-linked Immunosorbent Assay, Cell Culture, Confocal Microscopy, Microscopy

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Breaking the symmetry of cell contractility drives tubulogenesis via CXCL1 polarization.

doi: 10.1073/pnas.2315894121

Figure Lengend Snippet: Fig. 6. Suppressing the mechanosensing machinery CD29 abolishes CXCL1 expression and tubulogenesis. (A) Western blot analysis showing the expression of CD29 and tubulin-α in cells transfected with control (shCntl) or CD29-targeting shRNA (shCD29). (B) Tube formation on fish skin–derived collagen gel for 1 d in cells with shCntl or shCD29. ***P < 0.001. (C) Representative confocal microscopy images of cells with phalloidin (red) for microfilaments and DAPI (blue) for the nucleus on FITC-conjugated collagen gel for 1 d. (D) Analysis of Cxcl1 mRNA expression in cells grown on fish skin–derived collagen gel for 1 d. ***P < 0.001. (E) ELISA analysis of cell culture supernatant taken from cells grown on fish skin–derived collagen gel for 1 d. ***P < 0.001. (F) Flow chart shows breaking the symmetry of cell contractility by reducing matrix stiffness or myosin activity promotes CXCL1 polarization at cell protrusions. CXCL1–CXCR1 interaction at cell protrusions promotes cell polarization and ECM remodeling, which in turn, accelerates cell–cell connection and tube elongation.

Article Snippet: Immunofluorescent staining was performed using specific antibodies against CXCL1 (AF- 515- NA, R&D Systems), tubulin- α (05- 829, Merck), pMLC2 (95777, Cell Signaling Technology), and E- cadherin (610182, BD Biosciences) followed by Alexa Fluor- conjugated secondary antibodies (Invitrogen).

Techniques: Expressing, Western Blot, Transfection, Control, shRNA, Derivative Assay, Confocal Microscopy, Enzyme-linked Immunosorbent Assay, Cell Culture, Activity Assay