human itgb6 antibody  (R&D Systems)

Journal: Head & face medicine

Article Title: CRISPR/Cas9-mediated knock out of ITGB6 in human OSCC cells reduced migration and proliferation ability.

doi: 10.1186/s13005-024-00437-x

Figure Lengend Snippet: Fig. 1 Impact of ITGB6 mRNA expression on patient outcome. A Kaplan-Meier plot on the overall survival of patients afflicted with OSCC. Comparison of “high expression” (n = 83) mRNA ITGB6 levels and “low expression” (n = 82) mRNA ITGB6 levels. Significant longer overall survival of the “low expression” group, * p = 0.0355. B Plot of the mRNA expression levels of the group “low expression” and “high expression”. Significant difference in the mRNA expres sion levels of ITGB6, * p < 0.000001. C Comparison of perineural invasion in both groups. Significant increased perineural invasion in the “high expression” group, * p = 0.001214

Article Snippet: Subsequently the cells were stained with an APC-labeled anti-ITGB6 antibody (Miltenyi Biotec, Bergisch Gladbach, Germany; Catalog#: 130-111-454) for 30 min in phosphate-buffered saline (PBS) supplemented with 2% (v/v) FCS and 2 mM EDTA.

Techniques: Expressing, Comparison

Journal: Head & face medicine

Article Title: CRISPR/Cas9-mediated knock out of ITGB6 in human OSCC cells reduced migration and proliferation ability.

doi: 10.1186/s13005-024-00437-x

Figure Lengend Snippet: Fig. 2 CRISPR/Cas9-mediated knockout of ITGB6 in HN cells. A Sanger sequencing of HN WT (top) and HN ITGB6KO (bottom). Traces and sequences are presented. PAM sequence, gRNA binding and Cas9 cut site is marked. B Sequencing data of HN ITGB6KO analyzed by ICE online tool (Synthego). Decomposition of the found sequences with the individual contributions are presented (Sequence 1 1: 48%; Sequence 2: 41%). Sequences with contribu tions ≤ 2% are considered as background signal and are not shown. Sequence 1 shows an insertion of one base (marked in red color) (+ 1). Sequence 2 indicates a deletion of 1 base (-1) (deleted base shown in red color in the WT sequence). Overall, a strong indication of a heterozygous KO of ITGB6 with frameshift mutations on both alleles is presented. C Verification of ITGB6 gene KO of HN ITGB6KO on the protein level by FACS analysis. HN WT cells pres ent with a homogenous ITGB6 expression, whereas HN ITGB6KO indicates a loss of ITGB6 expression

Article Snippet: Subsequently the cells were stained with an APC-labeled anti-ITGB6 antibody (Miltenyi Biotec, Bergisch Gladbach, Germany; Catalog#: 130-111-454) for 30 min in phosphate-buffered saline (PBS) supplemented with 2% (v/v) FCS and 2 mM EDTA.

Techniques: CRISPR, Knock-Out, Sequencing, Binding Assay, Expressing

Journal: American Journal of Cancer Research

Article Title: Inhibition of ITGB6 stimulates potent anti-tumor responses in immunocompetent mouse models of head and neck squamous cell carcinoma and pancreatic adenocarcinoma

doi: 10.62347/MEAD1055

Figure Lengend Snippet: ITGB6 shows specificity and upregulation in human cancers. (A) Human Protein Atlas analysis of RNA expression of ITGB6 across various cancer types. Normalized RNA expression levels are given in fragments per kilobase million (FPKM). Data are acquired from TCGA. Graphs show the mean and SD. (B) Differential gene expression analysis of ITGB6 across tumor and normal tissue. Data are obtained from a standardized concatenated data set from GEO, GTex, TCGA, and TARGET databases. RNA-seq tumor and normal tissue samples are obtained from the same patients and from adjacent sites. Graphs were generated using the TNMplot tool and were normalized using DESeq2. Fold-change (FC) of medians and Mann-Whitney test p -values are shown.

Article Snippet: The human cell lines HCT116WT, Capan-2, FaDu, and CAL27 were screened via flow cytometry using an APC-conjugated human ITGB6 antibody (R&D Systems, # FAB4155A).

Techniques: RNA Expression, Gene Expression, RNA Sequencing, Generated, MANN-WHITNEY

Journal: American Journal of Cancer Research

Article Title: Inhibition of ITGB6 stimulates potent anti-tumor responses in immunocompetent mouse models of head and neck squamous cell carcinoma and pancreatic adenocarcinoma

doi: 10.62347/MEAD1055

Figure Lengend Snippet: ITGB6 knockout induces T-cell killing of HNSCC and PAAD cells. (A) ITGB6 RNA of selected human cancer cell lines from the Human Protein Atlas. (B) Flow cytometry analysis of the percentage of cancer cells that are ITGB6+ compared to unstained controls. (C) Western blot validation of the CRISPR knockout of ITGB6 in FaDu, CAL27, and Capan-2 cells. (D) Colony formation assay of CRISPR control (CTRL) cells and ITGB6 knockouts (KO-1) for FaDu and CAL27 cells (n = 3). (E) Quantification of colony formation assay. Statistical analysis was performed using an unpaired t-test. Randomly chosen fields of view from the co-culture of FaDu (F), CAL-27 (G), or Capan-2 (H) cells (green) and TALL-104 T-cells (blue) (n = 10). Cells were pretreated for 8 hrs with latent TGFβ. Quantification of cancer cell counts for FaDu (I), CAL27 (M), and Capan-2 (Q). Quantification of the percentage of cancer cells that are dead for FaDu (J), CAL27 (N), and Capan-2 (R). Quantification of TALL-104 T-cell counts for the FaDu cohort (K), the CAL27 cohort (O), and the Capan-2 cohort (S). Quantification of the percentage of T cells that are dead for the FaDu cohort (L), the CAL27 cohort (P), and the Capan-2 cohort (T). One-way ANOVA: P < 0.0001 (****), P < 0.01 (**).

Article Snippet: The human cell lines HCT116WT, Capan-2, FaDu, and CAL27 were screened via flow cytometry using an APC-conjugated human ITGB6 antibody (R&D Systems, # FAB4155A).

Techniques: Knock-Out, Flow Cytometry, Western Blot, Biomarker Discovery, CRISPR, Colony Assay, Control, Co-Culture Assay

Journal: American Journal of Cancer Research

Article Title: Inhibition of ITGB6 stimulates potent anti-tumor responses in immunocompetent mouse models of head and neck squamous cell carcinoma and pancreatic adenocarcinoma

doi: 10.62347/MEAD1055

Figure Lengend Snippet: Suppressed growth of ITGB6 -knockout tumors in immunocompetent mice. (A) Flow cytometry analysis of the percentage of mouse cancer cells that are ITGB6+ compared to unstained controls. (B) Western blot validation of CRISPR knockdown of ITGB6 in MOC1 and KPCY cells. (C) Colony formation assay of CRISPR control (CTRL) cells and ITGB6-knockout (KO-1, KO-2) cells for MOC1 and KPCY cells. (D) Quantification of colony formation assay. Statistical analysis was performed using one-way ANOVA: P < 0.05 (*). (E) Experimental timeline for C57BL/6 mice injected with syngeneic tumor cells. Tumor volume of mice starting upon tumor formation at day 5 for MOC1 (F) and KPCY (H) cohorts (n = 10). Mass of MOC1 (G) and KPCY (I) tumors harvested at day 25. Statistical comparison between CTRL and KO-1 cohorts was performed using an unpaired t-test: P < 0.001 (***), P < 0.05 (*). (J) Heat map of Luminex panel showing fold change of cytokines of tumor interstitial fluid harvested from MOC1 and KPCY tumors (n = 5). Red indicates upregulation and green downregulation of cytokines in ITGB6 KO tumors compared to control tumors. (K) Concentrations of select tumor interstitial fluid cytokines from MOC1 and KPCY (L) as measured by the Luminex panel. Statistical comparison between CTRL and KO-1 cohorts was performed using an unpaired t-test. Representative 10× images of MOC1 (M) and KPCY (O) from scanned IHC slides showing CD8 T-cell infiltration across control and ITGB6-deficient cells. Quantification of CD8-stained IHC slides for MOC1 (N) (control: n = 5, KO: n = 10) and KPCY (P) (control: n = 7, KO: n = 8) cohorts, showing p -values calculated using an unpaired t-test.

Article Snippet: The human cell lines HCT116WT, Capan-2, FaDu, and CAL27 were screened via flow cytometry using an APC-conjugated human ITGB6 antibody (R&D Systems, # FAB4155A).

Techniques: Knock-Out, Flow Cytometry, Western Blot, Biomarker Discovery, CRISPR, Knockdown, Colony Assay, Control, Injection, Comparison, Luminex, Staining

Journal: American Journal of Cancer Research

Article Title: Inhibition of ITGB6 stimulates potent anti-tumor responses in immunocompetent mouse models of head and neck squamous cell carcinoma and pancreatic adenocarcinoma

doi: 10.62347/MEAD1055

Figure Lengend Snippet: ITGB6 decreases overall survival in patients. Curves show the overall survival of patients with various cancer types stratified based on ITGB6 expression. Patients with high ITGB6 have expression levels above +0.25 σ of the mean, and ITGB6 low patients have expression below -0.25 σ. The graphs were generated using TCGA data through the cBioPortal. Statistical analysis of Kaplan-Meier curves and corresponding hazard ratios (HR) was performed using the Log-rank test.

Article Snippet: The human cell lines HCT116WT, Capan-2, FaDu, and CAL27 were screened via flow cytometry using an APC-conjugated human ITGB6 antibody (R&D Systems, # FAB4155A).

Techniques: Expressing, Generated

Journal: American Journal of Cancer Research

Article Title: Inhibition of ITGB6 stimulates potent anti-tumor responses in immunocompetent mouse models of head and neck squamous cell carcinoma and pancreatic adenocarcinoma

doi: 10.62347/MEAD1055

Figure Lengend Snippet: Immune checkpoint blockade is modulated by ITGB6 expression. (A) Pan-cancer analysis of ITGB6 RNA expression level normalized using DESeq2 of patients who are either responsive or unresponsive to immune checkpoint blockade. RNA data are extracted from a TCGA data set of multiple cancer types of patients who underwent immune checkpoint blockade therapy. Graphs were generated using the ROCplot tool, and the results of an unpaired t-test are shown. (B) Kaplan-Meier curves of overall survival of patients undergoing immune checkpoint blockade therapy stratified into low and high ITGB6 expression about the mean. Statistical analysis was performed using the Log-rank test.

Article Snippet: The human cell lines HCT116WT, Capan-2, FaDu, and CAL27 were screened via flow cytometry using an APC-conjugated human ITGB6 antibody (R&D Systems, # FAB4155A).

Techniques: Expressing, RNA Expression, Generated