Journal: Journal of Nuclear Medicine

Article Title: Clinical Translation of a 68 Ga-Labeled Integrin α v β 6 –Targeting Cyclic Radiotracer for PET Imaging of Pancreatic Cancer

doi: 10.2967/jnumed.119.237347

Figure Lengend Snippet: (A) Chemical structure of DOTA-cycratide. (B) Inhibition of 64Cu-cycratide binding to integrin αvβ6 on BxPC-3 cells by cycratide, DOTA-cycratide, and linear-pep. Data are shown as mean ± SD, n = 4. (C) Binding of 68Ga-cycratide to BxPC-3 with or without blocking of cold cycratide or linear-pep. %AD/106 cells = percentage of total added dose per million cells. Data are shown as mean ± SD, n = 4. (D) Metabolic stability of 68Ga-cycratide in blood and urine of BALB/c mice (data are representative of 3 independent experiments). **P < 0.01.

Article Snippet: After endogenous peroxidase activity had been abolished using 0.3% hydrogen peroxide and antigen had been retrieved by microwave, tumor tissues were incubated with rabbit antihuman integrin α v β 6 antibody (bs-5791r; Bioss) overnight at 4°C.

Techniques: Inhibition, Binding Assay, Blocking Assay

Journal: Journal of Nuclear Medicine

Article Title: Clinical Translation of a 68 Ga-Labeled Integrin α v β 6 –Targeting Cyclic Radiotracer for PET Imaging of Pancreatic Cancer

doi: 10.2967/jnumed.119.237347

Figure Lengend Snippet: (A) PET imaging of orthotopic pancreatic cancer lesions in nude mice at 0.5 h after injection of 68Ga-cycratide without or with blocking dose of cold cycratide. Tumors are indicated by arrows. (B) Hematoxylin and eosin (H&E) staining of tumor tissues harvested from orthotopic tumor model. (C) Left, immunofluorescence staining of integrin αvβ6 from tumor tissues harvested from orthotopic tumor model. Right, negative control with secondary antibody only.

Article Snippet: After endogenous peroxidase activity had been abolished using 0.3% hydrogen peroxide and antigen had been retrieved by microwave, tumor tissues were incubated with rabbit antihuman integrin α v β 6 antibody (bs-5791r; Bioss) overnight at 4°C.

Techniques: Imaging, Injection, Blocking Assay, Staining, Immunofluorescence, Negative Control

Journal: Journal of Nuclear Medicine

Article Title: Clinical Translation of a 68 Ga-Labeled Integrin α v β 6 –Targeting Cyclic Radiotracer for PET Imaging of Pancreatic Cancer

doi: 10.2967/jnumed.119.237347

Figure Lengend Snippet: (A) PET/CT images of female patient with suspected pancreatic cancer. Images were obtained at 1 h after intravenous administration of 68Ga-cycratide or 18F-FDG. Tumors are indicated by arrows. (B) Immunohistochemical (IHC) staining for integrin αvβ6 in tumor sample from same patient as in A. (C) Contrast-enhanced CT (CECT) image and PET/CT images of male patient with pancreatic cancer 7 mo after surgery at 1 h after administration of 68Ga-cycratide or 18F-FDG, as well as CECT image of same patient 3 mo later (10 mo after surgery). Occupancy lesions in CT are indicated by arrows.

Article Snippet: After endogenous peroxidase activity had been abolished using 0.3% hydrogen peroxide and antigen had been retrieved by microwave, tumor tissues were incubated with rabbit antihuman integrin α v β 6 antibody (bs-5791r; Bioss) overnight at 4°C.

Techniques: Positron Emission Tomography-Computed Tomography, Immunohistochemical staining, Immunohistochemistry

Journal: bioRxiv

Article Title: Cones and Cone Pathways Remain Functional in Advanced Retinal Degeneration

doi: 10.1101/2022.09.27.509752

Figure Lengend Snippet: (A and B) SIM images. (A) Ciliated cone photoreceptor cell. The cilium (arrow) is immunolabeled with acetylated tubulin (cyan). Inner segment (arrowhead) as well as the cilium is labeled with cone opsin antibodies (red). (B) Cone photoreceptor cell lacking a cilium. Inner segment only (arrowhead) is labeled by cone opsin antibodies (red). (C and D) Electron micrographs, a single cone photoreceptor has been colorized. (C) Ciliated cone photoreceptor with residual membrane amplifications (arrow). (D) Cone photoreceptor with inner segment (arrowhead) but lacking cilium. Scale bars (A) – (D), 1 μm.

Article Snippet: They were then washed with PBS, blocked with 5% donkey serum in PBS-0.2%TritonX100 (v/v) for 1h at room temperature (RT), and incubated with a mixture of 1:1000 rabbit anti-blue opsin antibody (AB5407, Millipore Sigma), 1:1000 rabbit anti-red/green opsin antibody (AB5405, Millipore Sigma), YF594-conjugated acetylated tubulin antibody (YF594-66200, Proteintech), and 2% donkey serum in PBS-0.2%TritonX100 (v/v) overnight at 4 °C.

Techniques: Immunolabeling, Labeling, Membrane

Journal: Advanced Science

Article Title: Inhibition of Integrin α v β 6 Activation of TGF‐ β Attenuates Tendinopathy

doi: 10.1002/advs.202104469

Figure Lengend Snippet: Integrin α v β 6 active latent TGF‐ β in response to mechanical stress drives the progression of tendinopathy. A) Representative images and B) quantification of immunostaining of tendon sections with antibodies against α v β 6 at 1, 4, and 8 weeks after DI or sham processing. Scale bar: 50 µm. All data are shown as the mean ± standard deviation ( n = 8 mice per group). * p < 0.05 compared with sham group as determined by one‐way analysis of variance. C) A schematic diagram of the ex vivo tensile model. D) Frequency distribution of tendon fiber orientation. E,G) Representative images and F,H) quantification of E,F) pSmad2 + cells and (G, H) TUNEL + cells. Scale bars: 50 µm. I) Western blot of pSmad2 and Smad2 levels in tendon lysates. J) Quantification of pSmad2 relative density relative to physiological loading group (20 g) in tendon lysates after above‐mentioned treatment. K) Quantitative analysis of maximum tensile force and stiffness of tendons. All data from B, F, H, J, and K are shown as the mean ± standard deviation ( n = 8 per group). * p < 0.05 compared with 200 g loaded group as determined by one‐way analysis of variance.

Article Snippet: Sections were incubated with primary antibodies to human CD68 (Abcam, Cambridge, UK, ab955, 1:200), human Col2 (Abcam, ab34712, 1:100), human MMP13 (Abcam, ab3208, 1:40), human/mouse pSmad2 (Santa Cruz Biotechnology, Dallas, TX, sc‐11769, 1:50), mouse Col2 (Abcam, ab185430, 1:100), mouse CD68 (Abcam, ab125212, 1:100), mouse MMP13 (Abcam, ab219620, 1:100), mouse green fluorescent protein (Abcam, ab290, 1:200), and integrin α v β 6 (Bioss Antibodies, Woburn, MA, bs‐5791R‐Biotin, 1:100) overnight at 4 °C.

Techniques: Immunostaining, Standard Deviation, Ex Vivo, TUNEL Assay, Western Blot

Journal: Communications Chemistry

Article Title: Paper-based electrochemical device for early detection of integrin αvβ6 expressing tumors

doi: 10.1038/s42004-024-01144-z

Figure Lengend Snippet: a Representative image of TRPS measurements showing particle size and concentration of S-EVs derived from DU145R80 (R80) cancer cell line (see text). b Upper panels: representative images of Transmission Electron micrograph (TEM) of S-EVs isolated from cell culture media. Magnification 49000X, scale bar 200 nm. The morphology is observed by negative staining. Lower panels: representative images of Immuno Electron micrograph (IEM) of S-EVs, labeled using antibodies specific to αvβ6 integrin and antibody binding was confirmed by Protein A gold-conjugate to 10 nm gold particles. Magnification 120000X, scale bars 100 nm. c Integrin β6 expression by ELISA expressed in terms of fold changes of S-EVs from PC3, A549 and HCT116 compared to R80-derived S-EVs. d Western blot analysis of S-EVs lysates from R80, PC3, HCT116 and A549 and tested with antibodies as indicated in the figure. Both TSG101 and CD81 were tested as positive EVs markers. β actin served as negative control. e Densitometry of Integrin αVβ6/CD81 expression in EVs, expressed as fold changes in either HEK293- or H460- S-EVs compared to PC3-derived S-EVs. Inset shows Western blot analysis of S-EVs lysates from PC3, HEK293 and H460 tested with antibodies as indicated in the figure. Both TSG101 and CD81 were tested as positive EVs markers. ɤ-tubulin served as negative control .

Article Snippet: For examination of immuno-gold labeled unfixed exosomes, rabbit anti-Integrin Alpha V + Beta 6 (1 μg/μL bs-5791R; Bioss Antibodies, Woburn, Massachusetts, USA) and Protein A gold-conjugate to 10 nm gold particles (PAG, purchased from Cell Microscopy Core Department of Cell Biology University Medical Center Utrecht, Utrecht, The Netherlands) were used.

Techniques: Concentration Assay, Derivative Assay, Transmission Assay, Isolation, Cell Culture, Negative Staining, Labeling, Binding Assay, Expressing, Enzyme-linked Immunosorbent Assay, Western Blot, Negative Control

Journal: Krimskii zhurnal eksperimental'noi i klinicheskoi meditsiny = Kryms'kyi zhurnal eksperymental'noi ta klinichnoi medytsyny = Crimean journal of experimental and clinical medicine

Article Title: THE IMPACT OF ALCOHOL ON PRO-METASTATIC N-GLYCOSYLATION IN PROSTATE CANCER

doi:

Figure Lengend Snippet: Schema of MGAT3 and MGAT5-mediated N-glycosylation. Bisecting GlcNAc blocks glycans branching via MGAT5.

Article Snippet: The primary antibodies used were: a) rabbit polyclonal – MGAT3 (Abcam, ab103427), MGAT5 (Abcam, ab68595), and ST14 (Matriptase) (Abcam, ab28266); b) mouse monoclonal – E-cadherin (Abcam, ab1416), β-actin (Sigma, A2228), integrin alpha V + beta 6 (Abcam, ab77906), and giantin (Abcam, ab37266); c) mouse polyclonal – GM130 (Abcam, ab169276).

Techniques:

Journal: Krimskii zhurnal eksperimental'noi i klinicheskoi meditsiny = Kryms'kyi zhurnal eksperymental'noi ta klinichnoi medytsyny = Crimean journal of experimental and clinical medicine

Article Title: THE IMPACT OF ALCOHOL ON PRO-METASTATIC N-GLYCOSYLATION IN PROSTATE CANCER

doi:

Figure Lengend Snippet: (A) MGAT3 and MGAT5 W-B of the cell lysate from LNCaP (c-26) and (c-86) cells; β-actin was used as a loading control. (B) Matriptase W-B of the plasma membrane (PM) fractions from LNCaP (c-26) and (c-86) cells; the PM samples were normalized by E-cadherin. (C) The L-PHA lectin W-B of Matriptase IP from LNCaP (c-26) and (c-86) cells. The bottom panel represents the input normalized for the Matriptase.

Article Snippet: The primary antibodies used were: a) rabbit polyclonal – MGAT3 (Abcam, ab103427), MGAT5 (Abcam, ab68595), and ST14 (Matriptase) (Abcam, ab28266); b) mouse monoclonal – E-cadherin (Abcam, ab1416), β-actin (Sigma, A2228), integrin alpha V + beta 6 (Abcam, ab77906), and giantin (Abcam, ab37266); c) mouse polyclonal – GM130 (Abcam, ab169276).

Techniques:

Journal: Krimskii zhurnal eksperimental'noi i klinicheskoi meditsiny = Kryms'kyi zhurnal eksperymental'noi ta klinichnoi medytsyny = Crimean journal of experimental and clinical medicine

Article Title: THE IMPACT OF ALCOHOL ON PRO-METASTATIC N-GLYCOSYLATION IN PROSTATE CANCER

doi:

Figure Lengend Snippet: Immunostaining of GM130 (green) and MGAT3 (red) (A), and giantin (green) and MGAT5 (red) (B) in the LNCaP cells: control and treated with 35 mM EtOH for 96 h. Nuclei were counterstained with DAPI (blue). All confocal images were acquired with the same imaging parameters; bars, 10 μm. (C) Quantification of Pearson’s overlap coefficient for indicated proteins for cells from A and B; means ± SD; * - p<0.001.

Article Snippet: The primary antibodies used were: a) rabbit polyclonal – MGAT3 (Abcam, ab103427), MGAT5 (Abcam, ab68595), and ST14 (Matriptase) (Abcam, ab28266); b) mouse monoclonal – E-cadherin (Abcam, ab1416), β-actin (Sigma, A2228), integrin alpha V + beta 6 (Abcam, ab77906), and giantin (Abcam, ab37266); c) mouse polyclonal – GM130 (Abcam, ab169276).

Techniques: Immunostaining, Imaging

Journal: Krimskii zhurnal eksperimental'noi i klinicheskoi meditsiny = Kryms'kyi zhurnal eksperymental'noi ta klinichnoi medytsyny = Crimean journal of experimental and clinical medicine

Article Title: THE IMPACT OF ALCOHOL ON PRO-METASTATIC N-GLYCOSYLATION IN PROSTATE CANCER

doi:

Figure Lengend Snippet: (A) MGAT3 and MGAT5 W-B of the lysate from LNCaP cells: control and treated with 35 mM EtOH for 96 h. (B) Integrin α5 W-B of the plasma membrane (PM) fraction from the cells presented in A; the PM samples were normalized by E-cadherin

Article Snippet: The primary antibodies used were: a) rabbit polyclonal – MGAT3 (Abcam, ab103427), MGAT5 (Abcam, ab68595), and ST14 (Matriptase) (Abcam, ab28266); b) mouse monoclonal – E-cadherin (Abcam, ab1416), β-actin (Sigma, A2228), integrin alpha V + beta 6 (Abcam, ab77906), and giantin (Abcam, ab37266); c) mouse polyclonal – GM130 (Abcam, ab169276).

Techniques:

Journal: Krimskii zhurnal eksperimental'noi i klinicheskoi meditsiny = Kryms'kyi zhurnal eksperymental'noi ta klinichnoi medytsyny = Crimean journal of experimental and clinical medicine

Article Title: THE IMPACT OF ALCOHOL ON PRO-METASTATIC N-GLYCOSYLATION IN PROSTATE CANCER

doi:

Figure Lengend Snippet: (A) 3D SIM reconstruction of giantin (green) and MGAT3 (red) IF in normal prostate and tissue section of PCa patients (Gleason 7); bars, 5 µm. White boxes indicate area enlarged below. (B) Quantification of colocalization of giantin and MGAT3 from the sections in A; 10 patients for each group were observed; means ± SD; * - p<0.001.

Article Snippet: The primary antibodies used were: a) rabbit polyclonal – MGAT3 (Abcam, ab103427), MGAT5 (Abcam, ab68595), and ST14 (Matriptase) (Abcam, ab28266); b) mouse monoclonal – E-cadherin (Abcam, ab1416), β-actin (Sigma, A2228), integrin alpha V + beta 6 (Abcam, ab77906), and giantin (Abcam, ab37266); c) mouse polyclonal – GM130 (Abcam, ab169276).

Techniques:

Journal: Krimskii zhurnal eksperimental'noi i klinicheskoi meditsiny = Kryms'kyi zhurnal eksperymental'noi ta klinichnoi medytsyny = Crimean journal of experimental and clinical medicine

Article Title: THE IMPACT OF ALCOHOL ON PRO-METASTATIC N-GLYCOSYLATION IN PROSTATE CANCER

doi:

Figure Lengend Snippet: The working model of alcohol effect on N-glycosylation in PCa. In low aggressive PCa cells, pro-metastatic proteins are modified by MGAT3, which reduces their binding to the Gal-3, thus blocking their stabilization and retention at the cell surface. In alcohol-treated PCa cells, Golgi is fragmented and domination of MGAT5-mediated glycosylation promotes plasma membrane expression of proteins via their strong interaction with Gal-3. This, in turn, facilitates PCa progression.

Article Snippet: The primary antibodies used were: a) rabbit polyclonal – MGAT3 (Abcam, ab103427), MGAT5 (Abcam, ab68595), and ST14 (Matriptase) (Abcam, ab28266); b) mouse monoclonal – E-cadherin (Abcam, ab1416), β-actin (Sigma, A2228), integrin alpha V + beta 6 (Abcam, ab77906), and giantin (Abcam, ab37266); c) mouse polyclonal – GM130 (Abcam, ab169276).

Techniques: Modification, Binding Assay, Blocking Assay, Expressing

Journal: Krimskii zhurnal eksperimental'noi i klinicheskoi meditsiny = Kryms'kyi zhurnal eksperymental'noi ta klinichnoi medytsyny = Crimean journal of experimental and clinical medicine

Article Title: THE IMPACT OF ALCOHOL ON PRO-METASTATIC N-GLYCOSYLATION IN PROSTATE CANCER

doi:

Figure Lengend Snippet: (A) MGAT3 and MGAT5 W-B of the cell lysate from LNCaP (c-26) and (c-86) cells; β-actin was used as a loading control. (B) Matriptase W-B of the plasma membrane (PM) fractions from LNCaP (c-26) and (c-86) cells; the PM samples were normalized by E-cadherin. (C) The L-PHA lectin W-B of Matriptase IP from LNCaP (c-26) and (c-86) cells. The bottom panel represents the input normalized for the Matriptase.

Article Snippet: The primary antibodies used were: a) rabbit polyclonal – MGAT3 (Abcam, ab103427), MGAT5 (Abcam, ab68595), and ST14 (Matriptase) (Abcam, ab28266); b) mouse monoclonal – E-cadherin (Abcam, ab1416), β-actin (Sigma, A2228), integrin alpha V + beta 6 (Abcam, ab77906), and giantin (Abcam, ab37266); c) mouse polyclonal – GM130 (Abcam, ab169276).

Techniques:

Journal: Krimskii zhurnal eksperimental'noi i klinicheskoi meditsiny = Kryms'kyi zhurnal eksperymental'noi ta klinichnoi medytsyny = Crimean journal of experimental and clinical medicine

Article Title: THE IMPACT OF ALCOHOL ON PRO-METASTATIC N-GLYCOSYLATION IN PROSTATE CANCER

doi:

Figure Lengend Snippet: (A) MGAT5 W-B of the cell lysate from LNCaP (c-26) treated with a scramble of MGAT5 siRNAs; β-actin was used as a loading control. (B) Matriptase W-B of the plasma membrane (PM) fraction from cells in A; the PM samples were normalized by E-cadherin.

Article Snippet: The primary antibodies used were: a) rabbit polyclonal – MGAT3 (Abcam, ab103427), MGAT5 (Abcam, ab68595), and ST14 (Matriptase) (Abcam, ab28266); b) mouse monoclonal – E-cadherin (Abcam, ab1416), β-actin (Sigma, A2228), integrin alpha V + beta 6 (Abcam, ab77906), and giantin (Abcam, ab37266); c) mouse polyclonal – GM130 (Abcam, ab169276).

Techniques:

Journal: Krimskii zhurnal eksperimental'noi i klinicheskoi meditsiny = Kryms'kyi zhurnal eksperymental'noi ta klinichnoi medytsyny = Crimean journal of experimental and clinical medicine

Article Title: THE IMPACT OF ALCOHOL ON PRO-METASTATIC N-GLYCOSYLATION IN PROSTATE CANCER

doi:

Figure Lengend Snippet: (A) MGAT3 and MGAT5 W-B of the lysate from LNCaP cells: control and treated with 35 mM EtOH for 96 h. (B) Integrin α5 W-B of the plasma membrane (PM) fraction from the cells presented in A; the PM samples were normalized by E-cadherin

Article Snippet: The primary antibodies used were: a) rabbit polyclonal – MGAT3 (Abcam, ab103427), MGAT5 (Abcam, ab68595), and ST14 (Matriptase) (Abcam, ab28266); b) mouse monoclonal – E-cadherin (Abcam, ab1416), β-actin (Sigma, A2228), integrin alpha V + beta 6 (Abcam, ab77906), and giantin (Abcam, ab37266); c) mouse polyclonal – GM130 (Abcam, ab169276).

Techniques: