Journal: Clinical and Translational Gastroenterology

Article Title: β 6 -Integrin Serves as a Potential Serum Marker for Diagnosis and Prognosis of Pancreatic Adenocarcinoma

doi: 10.14309/ctg.0000000000000395

Figure Lengend Snippet: Validation of ITGB6 as possible biomarker for PAC. ( a ) Serum ITGB6 levels were assessed in a prospective study cohort of 27 patients with PAC (cohort 1, N = 27). As control served 9 healthy volunteers (Ctrl; N = 9) and 10 patients with cP (N = 10). Significant differences in ITGB6 levels were observed between patients with PAC and Ctrl ( P = 0.019). ( b ) Comparing patients with or without distant metastatic PAC, serum ITGB6 levels were not significantly different. However, a significant increase in ITGB6 levels was observed between Ctrl and patients with nonmetastatic PAC ( P = 0.019). ( c ) To assess the prognostic value of serum ITGB6 levels, patients with PAC were plotted against their status of survival at time of blood assessment. A significant difference in ITGB6 concentration was observed between patients with PAC with status alive vs dead ( P = 0.007). ( d ) Two-dimensional scatterplots depict serum ITGB6 levels in relation to the serum CA19-9 levels from each cP ( r s = 0.511; P = 0.132) and patient with PAC ( r s = 0.210; P = 0.302). In ( a – c ), Mann–Whitney U Exact and Sig. 2-Tailed test were performed, and in ( d ), Spearman's rho correlation ( r s ) and Sig. 2-tailed test were performed. ( d ) Red lines indicate ITGB6 cutoff value at 0.1 ng/mL and black line CA19-9 cutoff value at 37.0 kU/L, respectively. cP, chronic pancreatitis; Ctrl, control; ITGB6, β 6 -integrin; PAC, pancreatic adenocarcinoma; r s, Spearman's rho correlation. *Means P < 0.05.

Article Snippet: A tissue microarray (TMA) with formalin-fixed paraffin-embedded PAC tissue specimens from 83 patients was automatically stained with a human anti-ITGB6 polyclonal antibody raised in rabbit (HPA023626, dilution 1:100; Atlas Antibodies AB, Bromma, Sweden) using a validated standard protocol on a Ventana automat (Ventana Medical System, Tucson, AZ).

Techniques: Biomarker Discovery, Control, Concentration Assay, MANN-WHITNEY

Journal: Clinical and Translational Gastroenterology

Article Title: β 6 -Integrin Serves as a Potential Serum Marker for Diagnosis and Prognosis of Pancreatic Adenocarcinoma

doi: 10.14309/ctg.0000000000000395

Figure Lengend Snippet: Serum ITGB6 levels may predict overall and progression-free survival in patients with PAC. ( a ) Survival rates and ( b ) progression-free survival of patients with PAC who underwent systemic therapy are plotted according to serum ITGB6 changes after initiation (N = 24; cohort 2). ( c ) Two-dimensional scatterplots depict lymph node ratio (LNR) in relation to exits after surgery (months) for each patient in accordance to the ITGB6 protein expression pattern. Hazard ratio (HR) was calculated with the Cox regression. ( d ) Overall survival in AU12 patients with PAC (N582; cohort 3) was plotted according to ITGB6 protein expression pattern. Kaplan-Meier estimation was performed and plotted accordingly. The log-rank P values are indicated. ITGB6, β 6 -integrin; PAC, pancreatic adenocarcinoma.

Article Snippet: A tissue microarray (TMA) with formalin-fixed paraffin-embedded PAC tissue specimens from 83 patients was automatically stained with a human anti-ITGB6 polyclonal antibody raised in rabbit (HPA023626, dilution 1:100; Atlas Antibodies AB, Bromma, Sweden) using a validated standard protocol on a Ventana automat (Ventana Medical System, Tucson, AZ).

Techniques: Expressing

Journal: Clinical and Translational Gastroenterology

Article Title: β 6 -Integrin Serves as a Potential Serum Marker for Diagnosis and Prognosis of Pancreatic Adenocarcinoma

doi: 10.14309/ctg.0000000000000395

Figure Lengend Snippet: ITGB6 protein expression in PAC tissue specimens by IHC. Representative TMA specimens (from cohort 3, N = 83) illustrate ITGB6-negative (no [0] or mild [1+] expression) and ITGB6-positive PAC specimens (moderate [2+] or strong [3+] expression; scale bars: upper panel 200 μm; lower panel 100 μm). IHC, immunohistochemistry; ITGB6, β 6 -integrin; PAC, pancreatic adenocarcinoma; TMA, tissue microarray.

Article Snippet: A tissue microarray (TMA) with formalin-fixed paraffin-embedded PAC tissue specimens from 83 patients was automatically stained with a human anti-ITGB6 polyclonal antibody raised in rabbit (HPA023626, dilution 1:100; Atlas Antibodies AB, Bromma, Sweden) using a validated standard protocol on a Ventana automat (Ventana Medical System, Tucson, AZ).

Techniques: Expressing, Immunohistochemistry, Microarray

Journal: Cell & Bioscience

Article Title: Protein expression of eIF4E and integrin αvβ6 in colon cancer can predict clinical significance, reveal their correlation and imply possible mechanism of interaction

doi: 10.1186/2045-3701-4-23

Figure Lengend Snippet: Association between eIF4E expression, integrin αvβ6 expression and clinicopathologic variables in colon cancer cases

Article Snippet: As primary antibodies, prediluted anti-Integrin β6 polyclonal rabbit antibody (1:200; Proteintech Group, Inc., Cat.No.19695-1-AP, Chicago, USA), anti- eIF-4E rabbit monoclonal antibody (1:100; Epitomics, Inc., Cat.No.

Techniques: Expressing

Journal: Cell & Bioscience

Article Title: Protein expression of eIF4E and integrin αvβ6 in colon cancer can predict clinical significance, reveal their correlation and imply possible mechanism of interaction

doi: 10.1186/2045-3701-4-23

Figure Lengend Snippet: Integrin β6 expression in human colonic cancer. First row: (a) , (b) , (c) (Scale bar 200 μm.); second row: (d) , (e) , (f) , corresponding to each figure above (Scale bar 100 μm). (a) and (d) was obtained from paracancerous normal tissue of patient with colon cancer, with negative staining. (b) and (e) was obtained from colonic carcinoma tissue,with negative Integrin β6 expression. (c) and (f) was obtained from colonic carcinoma, with positive Integrin β6 expression.

Article Snippet: As primary antibodies, prediluted anti-Integrin β6 polyclonal rabbit antibody (1:200; Proteintech Group, Inc., Cat.No.19695-1-AP, Chicago, USA), anti- eIF-4E rabbit monoclonal antibody (1:100; Epitomics, Inc., Cat.No.

Techniques: Expressing, Negative Staining

Journal: Cell & Bioscience

Article Title: Protein expression of eIF4E and integrin αvβ6 in colon cancer can predict clinical significance, reveal their correlation and imply possible mechanism of interaction

doi: 10.1186/2045-3701-4-23

Figure Lengend Snippet: Correlation between integrin αvβ6 expression and eIF4E expression in human colonic carcinoma tissues (r = 0.299, P < 0.001)

Article Snippet: As primary antibodies, prediluted anti-Integrin β6 polyclonal rabbit antibody (1:200; Proteintech Group, Inc., Cat.No.19695-1-AP, Chicago, USA), anti- eIF-4E rabbit monoclonal antibody (1:100; Epitomics, Inc., Cat.No.

Techniques: Expressing

Journal: Cell & Bioscience

Article Title: Protein expression of eIF4E and integrin αvβ6 in colon cancer can predict clinical significance, reveal their correlation and imply possible mechanism of interaction

doi: 10.1186/2045-3701-4-23

Figure Lengend Snippet: Overall survival according to eIF4E with integrin αvβ6 expression (P = 0.028, The log-rank test).

Article Snippet: As primary antibodies, prediluted anti-Integrin β6 polyclonal rabbit antibody (1:200; Proteintech Group, Inc., Cat.No.19695-1-AP, Chicago, USA), anti- eIF-4E rabbit monoclonal antibody (1:100; Epitomics, Inc., Cat.No.

Techniques: Expressing

Journal: The Journal of Clinical Investigation

Article Title: Immunoglobulin light chains generate proinflammatory and profibrotic kidney injury

doi: 10.1172/JCI125517

Figure Lengend Snippet: (A and B) Flow cytometric studies demonstrated that both the κ2 and λ3 FLCs increased the number of cells expressing ITGB6 (n = 4 experiments in each group). Pretreatment of HK-2 cells with ITGB6 shRNA prevented this response. (C) Along with the changes in surface localization of ITGB6, pretreatment of the cells with ITGB6 shRNA also inhibited the production of active TGF-β (n = 4 experiments in each group). Data are expressed as the mean ± SEM. *P < 0.0009 compared with the other 2 corresponding groups (ANOVA).

Article Snippet: Additional antibodies obtained commercially included those directed against SMAD2/3 (C4T) (MilliporeSigma); κ FLC (no. A0100, Dako, Agilent Technologies); ITGB6 (catalog TA321227, OriGene Technologies); GAPDH (catalog Ab8245, Abcam); HO-1 (ADI-SPA-894-F; Enzo Life Sciences); rat F4/80 (catalog 14480182, Invitrogen, Thermo Fisher Scientific); IL-1β (catalog Ab9722, Abcam); NIMP-R14 (sc-59338, Santa Cruz Biotechnology); and KIM-1 (AF1817, R&D Systems).

Techniques: Expressing, shRNA

Journal: The Journal of Clinical Investigation

Article Title: Immunoglobulin light chains generate proinflammatory and profibrotic kidney injury

doi: 10.1172/JCI125517

Figure Lengend Snippet: (A) Neutrophils, detected using anti–NIMP-R14 antibodies (red fluorescence), were present in greater numbers (white arrowheads) only in the Stat1+/+ mice treated with the higher dose of κ2 FLC. Proximal tubule expression of ITGB6 (green fluorescence) was readily detected. The IgG controls (left panel) showed minimal background staining of the samples. (B) M2 macrophage numbers, detected by colocalization of F4/80 (red fluorescence) with HO-1 (green fluorescence), were increased (white arrowheads) only in the Stat1+/+ mice treated with κ2 FLC. The inset shows a magnified image of a F4/80+HO-1+ cell (colocalized stain is shown in yellow). The IgG controls (left panel) showed minimal background staining of the samples. T, tubule. n = 8–10 mice/group. Data are expressed as the mean ± SEM of cells per HPF. *P < 0.0001 compared with the other 3 groups (ANOVA). Scale bars: 50 μm. Original magnification ×1200.

Article Snippet: Additional antibodies obtained commercially included those directed against SMAD2/3 (C4T) (MilliporeSigma); κ FLC (no. A0100, Dako, Agilent Technologies); ITGB6 (catalog TA321227, OriGene Technologies); GAPDH (catalog Ab8245, Abcam); HO-1 (ADI-SPA-894-F; Enzo Life Sciences); rat F4/80 (catalog 14480182, Invitrogen, Thermo Fisher Scientific); IL-1β (catalog Ab9722, Abcam); NIMP-R14 (sc-59338, Santa Cruz Biotechnology); and KIM-1 (AF1817, R&D Systems).

Techniques: Fluorescence, Expressing, Staining

Journal: The Journal of Clinical Investigation

Article Title: Immunoglobulin light chains generate proinflammatory and profibrotic kidney injury

doi: 10.1172/JCI125517

Figure Lengend Snippet: (A) Both doses of the κ2 FLCs prompted increases in ITGB6 levels of cortical lysates from Stat1+/+ mice (n = 6 mice/group). Data are expressed as the mean ± SEM. **P ≤ 0.006 compared with each of the other 4 groups (ANOVA). Relative ITGB6 levels were greater (P = 0.0096, ANOVA) in the PBS-treated Stat1+/+ mice compared with levels in the PBS-treated Stat1–/– mice. (B) A dose-dependent effect of κ2 FLC on p-SMAD2/3 was observed in Stat1+/+ mice but produced no changes in p-SMAD2/3 in Stat1–/– mice. Because SMAD2/3 levels were remarkably low in the kidney cortex of Stat1–/– mice, the data were factored by the density of GAPDH in the samples. n = 6 mice/group. Data are expressed as the mean ± SEM. *P < 0.005 compared with each of the other 5 groups in the experiment (ANOVA).

Article Snippet: Additional antibodies obtained commercially included those directed against SMAD2/3 (C4T) (MilliporeSigma); κ FLC (no. A0100, Dako, Agilent Technologies); ITGB6 (catalog TA321227, OriGene Technologies); GAPDH (catalog Ab8245, Abcam); HO-1 (ADI-SPA-894-F; Enzo Life Sciences); rat F4/80 (catalog 14480182, Invitrogen, Thermo Fisher Scientific); IL-1β (catalog Ab9722, Abcam); NIMP-R14 (sc-59338, Santa Cruz Biotechnology); and KIM-1 (AF1817, R&D Systems).

Techniques: Produced

Journal: Journal of Neurotrauma

Article Title: MicroRNA-711–Induced Downregulation of Angiopoietin-1 Mediates Neuronal Cell Death

doi: 10.1089/neu.2017.5572

Figure Lengend Snippet: Ang-1 activates the Akt pathway in neurons through both Tie-2 and β1-integrin signaling. (A) RCNs were treated with Ang-1 for the indicated time. Whole-cell lysates were fractioned on SDS-polyacrylamide gel and immunoblotted with antibodies against Tie-2, phosphor-Tie2 (Y992), β1-integrin, FAK, phosphor-FAK (Tyr397), phospho-Akt (Ser473), phosphor-GSK3α/β, and β-actin. (B) Protein levels were quantified by densitometry, normalized to β-actin, and presented as fold change compared to control untreated levels. Data represent the mean ± SEM. One-way ANOVA, SNK post-hoc analysis; *p < 0.05; **p < 0.01; ***p < 0.001 versus cnt; ^^^p < 0.001 versus etoposide treated (N = 3). Akt, protein kinase B; Ang-1, angiopoietin-1; ANOVA, analysis of variance; FAK, focal adhesion kinase; GSK3α/β, glycogen synthase kinase 3α/β; RCNs, rat cortical neurons; SDS, sodium dodecyl sulfate; SEM, standard error of the mean; SNK, Student–Newman–Keuls; Tie-2, Tunica interna endothelial cell kinase 2.

Article Snippet: Antibodies The following antibodies were used in this study: Histone H2A.X (ab11175; Abcam); apoptosis-inducing factor (AIF; sc-13116); cytochrome c (sc-13560) and Bim (sc-11425; Santa Cruz Biotechnology, Santa Cruz, CA); cleaved caspase-3 (#9661), cleaved PARP (poly (ADP-ribose) polymerase-1; #9545); phospho-GSK3α/β (glycogen synthase kinase 3α/β; Ser21/9; #9331); forkhead box O3a (FoxO3a; 75D8; #2497); Akt (pan; 11E7; #4685); FAK (#3285); phospho-FAK (Tyr397; #3283); phospho-p53 (Ser15; #9284); p53 (#2524); mitogen-activated protein kinase kinase 1 and 2 (MEK1/2; #8727); Lamin A/C (#4777); cyclooxegenase IV (COX-IV; #4844); phospho-Akt (Ser473; #4060; Cell Signaling Technology; GAPDH (ADI-CSA-335) and α-fodrin (BML-FG6090); active Bax (ALX-804-224-C100; Enzo Life Sciences, Inc., Farmingdale, NY); PUMA (#3041), Noxa (#2437; ProSci Incorporated, Poway, CA); β-actin (A1978); MAP2 (M3696; Sigma-Aldrich); integrin β1 (MAB1965); GFAP (MAB360; Millipore, Billerica, MA); Ang-1 (GTX28451; GeneTex Inc., Irvine, CA); Tie-2 (#AF762); phospho-Tie-2 (#AF2720; R&D Systems); and ionized calcium-binding adaptor molecule 1 (019-19741; Wako Pure Chemical Industries, Osaka, Japan).

Techniques:

Journal: Journal of Neurotrauma

Article Title: MicroRNA-711–Induced Downregulation of Angiopoietin-1 Mediates Neuronal Cell Death

doi: 10.1089/neu.2017.5572

Figure Lengend Snippet: Blocking of Tie-2 and β1-integrin signaling inhibits the neuroprotective effect of Ang-1. RCNs were treated for 30 min with antibodies against Tie2 (50 μg/mL; A); antibodies against β1-integrin (5 μg/mL; B); combination of Tie2 (50 μg/mL) and β1-integrin (5 μg/mL; C) and FAK inhibitor PF573228 (100 nM; D). Next RCNs were treated with Ang-1 and etoposide (Etop) or etoposide alone. Twenty-four hours later, LDH release was measured. Data are expressed as percentage of control untreated neurons. (E) RCNs were treated with etoposide alone; Ang-1 and etoposide and etoposide; and Ang-1 etoposide and Akt inhibitor (2.875 μM). Cell death and cell viability were measured using the LDH and Calcein AM (calcein-acetoxymethyl ester). Data represent the mean ± SEM. One-way ANOVA, SNK post-hoc analysis; *p < 0.05; **p < 0.01; ***p < 0.001 versus cnt; ^^p < 0.01; ^^^p < 0.001 versus etoposide treated; #p < 0.05; ##p < 0.01; ###p < 0.001 versus Ang-1 + etoposide treated; ∼∼p < 0.01; ∼∼∼p < 0.001 versus Tie-2 blocking/Ang-1/etoposide treated; &&p < 0.01 versus β1-integrin blocking/Ang-1/etoposide treated (N = 3). Akt, protein kinase B; Ang-1, angiopoietin-1; ANOVA, analysis of variance; FAK, focal adhesion kinase; LDH, lactate dehydrogenase; RCNs, rat cortical neurons; SEM, standard error of the mean; SNK, Student–Newman–Keuls; Tie-2, Tunica interna endothelial cell kinase 2.

Article Snippet: Antibodies The following antibodies were used in this study: Histone H2A.X (ab11175; Abcam); apoptosis-inducing factor (AIF; sc-13116); cytochrome c (sc-13560) and Bim (sc-11425; Santa Cruz Biotechnology, Santa Cruz, CA); cleaved caspase-3 (#9661), cleaved PARP (poly (ADP-ribose) polymerase-1; #9545); phospho-GSK3α/β (glycogen synthase kinase 3α/β; Ser21/9; #9331); forkhead box O3a (FoxO3a; 75D8; #2497); Akt (pan; 11E7; #4685); FAK (#3285); phospho-FAK (Tyr397; #3283); phospho-p53 (Ser15; #9284); p53 (#2524); mitogen-activated protein kinase kinase 1 and 2 (MEK1/2; #8727); Lamin A/C (#4777); cyclooxegenase IV (COX-IV; #4844); phospho-Akt (Ser473; #4060; Cell Signaling Technology; GAPDH (ADI-CSA-335) and α-fodrin (BML-FG6090); active Bax (ALX-804-224-C100; Enzo Life Sciences, Inc., Farmingdale, NY); PUMA (#3041), Noxa (#2437; ProSci Incorporated, Poway, CA); β-actin (A1978); MAP2 (M3696; Sigma-Aldrich); integrin β1 (MAB1965); GFAP (MAB360; Millipore, Billerica, MA); Ang-1 (GTX28451; GeneTex Inc., Irvine, CA); Tie-2 (#AF762); phospho-Tie-2 (#AF2720; R&D Systems); and ionized calcium-binding adaptor molecule 1 (019-19741; Wako Pure Chemical Industries, Osaka, Japan).

Techniques: Blocking Assay

Journal: Cell Death & Disease

Article Title: Myofibroblast induces hepatocyte-to-ductal metaplasia via laminin–ɑvβ6 integrin in liver fibrosis

doi: 10.1038/s41419-020-2372-9

Figure Lengend Snippet: Key resourse table.

Article Snippet: Rabbit anti-ITGB6 , ABclonal technology , WB , A16904.

Techniques: Immunohistochemistry-IF, Flow Cytometry, Recombinant, Solvent, Cell Culture, Staining, RNA Extraction, Isolation, Transfection, Software, Imaging