Journal: American Journal of Translational Research

Article Title: Lyn kinase enhanced hepatic fibrosis by modulating the activation of hepatic stellate cells

doi:

Figure Lengend Snippet: Increased activation of HSCs in Lyn TG mice upon CCl4 treatment. A: Representative immunofluorescence photomicrographs of HSC activation marker (Αvβ3, α-SMA and desmin) expression in liver tissues from WT mice and Lyn TG mice after administration of CCl4 (×200 magnification). B: The fluorescence intensity of desmin in 8 random fields. C: The fluorescence intensity of α-SMA in 8 random fields. D: The fluorescence intensity of Αvβ3 in 8 random fields. E: TGF-β1 levels in the liver tissues were determined by ELISA. All of the data are presented as the mean ± s.d. One-way ANOVA (Tukey-Kramer post-test) was used for Statistical analysis.

Article Snippet: After blocking with 1% BSA (Sigma-Aldrich) containing 0.05% Tween 20 for 1 hour, the specimens were then incubated with anti-Lyn antibody (Santa Cruz, CA, sc-15, AB_2281450), anti-integrin αvβ3 antibody (Bioss, China, bs-1310R), anti-α-SMA antibody (Bioss, China, bs-0189R), anti-desmin antibody (Bioss, China, bs-1026R), and anti-COL1α1 antibody (Bioss, China, bs-10423R).

Techniques: Activation Assay, Immunofluorescence, Marker, Expressing, Fluorescence, Enzyme-linked Immunosorbent Assay

Journal: American Journal of Translational Research

Article Title: Lyn kinase enhanced hepatic fibrosis by modulating the activation of hepatic stellate cells

doi:

Figure Lengend Snippet: Lyn kinase increased the activation of HSCs in vitro. Immunohistochemical staining for HSC activation marker (Αvβ3, α-SMA and COL1α1) expression in TGF-β1-induced LX-2 and Lyn+/+ LX-2. A: Representative photomicrographs and fluorescence intensity of α-SMA in cells (×400 magnification). B: Representative photomicrographs and fluorescence intensity of Αvβ3 in cells (×400 magnification). C: Representative photomicrographs and fluorescence intensity of COL1α1 in cells (×400 magnification). All of the data are presented as the mean ± s.d. One-way ANOVA (Tukey-Kramer post-test) was used for Statistical analysis. LX-2 cell lines transfected with lentiviral vector expressing Lyn were described as Lyn+/+ cells. LX-2 cell lines transfected with control lentiviral vector were described as WT cells.

Article Snippet: After blocking with 1% BSA (Sigma-Aldrich) containing 0.05% Tween 20 for 1 hour, the specimens were then incubated with anti-Lyn antibody (Santa Cruz, CA, sc-15, AB_2281450), anti-integrin αvβ3 antibody (Bioss, China, bs-1310R), anti-α-SMA antibody (Bioss, China, bs-0189R), anti-desmin antibody (Bioss, China, bs-1026R), and anti-COL1α1 antibody (Bioss, China, bs-10423R).

Techniques: Activation Assay, In Vitro, Immunohistochemical staining, Staining, Marker, Expressing, Fluorescence, Transfection, Plasmid Preparation

Journal: American Journal of Translational Research

Article Title: Lyn kinase enhanced hepatic fibrosis by modulating the activation of hepatic stellate cells

doi:

Figure Lengend Snippet: The Src-specific inhibitor PP2 suppressed HSC activation. A: Immunohistochemical staining for HSC activation marker (Αvβ3) expression in TGF-β1-induced LX-2 and Lyn+/+ LX-2 in the presence of PP2 (×400 magnification). The fluorescence intensity of Αvβ3 in cells was measured. B: Apoptotic cells of LX-2 were detected by flow cytometry. All of the data are presented as the mean ± s.d. One-way ANOVA (Tukey-Kramer post-test) was used for Statistical analysis.

Article Snippet: After blocking with 1% BSA (Sigma-Aldrich) containing 0.05% Tween 20 for 1 hour, the specimens were then incubated with anti-Lyn antibody (Santa Cruz, CA, sc-15, AB_2281450), anti-integrin αvβ3 antibody (Bioss, China, bs-1310R), anti-α-SMA antibody (Bioss, China, bs-0189R), anti-desmin antibody (Bioss, China, bs-1026R), and anti-COL1α1 antibody (Bioss, China, bs-10423R).

Techniques: Activation Assay, Immunohistochemical staining, Staining, Marker, Expressing, Fluorescence, Flow Cytometry

Journal: BMC Women's Health

Article Title: AlphaVBeta3 Integrin expression within uterine endometrium in unexplained infertility: a prospective cohort study

doi: 10.1186/s12905-017-0438-3

Figure Lengend Snippet: Immunostained slide with anti-avβ3 integrin, DAB chromogen, magnification x 200, score 0 in a case of unexplained infertility group

Article Snippet: After blocking with non-immune 4% goat serum incubated with tissues for 30 min, the primary antibody using monoclonal mouse IgG antibodies directed against human αvβ3 integrin (Human integrin alphaV beta3 MAb (clone 23C6)®, MAB3050, R&D systems, Minneapolis, USA) was added to these sections.

Techniques:

Journal: BMC Women's Health

Article Title: AlphaVBeta3 Integrin expression within uterine endometrium in unexplained infertility: a prospective cohort study

doi: 10.1186/s12905-017-0438-3

Figure Lengend Snippet: Immunostained slide with anti-avβ3 integrin, DAB chromogen, magnification x 200, score 3 in a case of fertility group. Reactivity is mainly in the galndular epithelium)

Article Snippet: After blocking with non-immune 4% goat serum incubated with tissues for 30 min, the primary antibody using monoclonal mouse IgG antibodies directed against human αvβ3 integrin (Human integrin alphaV beta3 MAb (clone 23C6)®, MAB3050, R&D systems, Minneapolis, USA) was added to these sections.

Techniques:

Journal: BMC Women's Health

Article Title: AlphaVBeta3 Integrin expression within uterine endometrium in unexplained infertility: a prospective cohort study

doi: 10.1186/s12905-017-0438-3

Figure Lengend Snippet: Immunostained slide with anti- vβ3 integrin, DAB chromogen, magnification x 40 (Low Power), score 3 in A case of fertility group. Reactivity could be seen in both Luminal Epithelium (LE) the Glandular Epithelium (GE)

Article Snippet: After blocking with non-immune 4% goat serum incubated with tissues for 30 min, the primary antibody using monoclonal mouse IgG antibodies directed against human αvβ3 integrin (Human integrin alphaV beta3 MAb (clone 23C6)®, MAB3050, R&D systems, Minneapolis, USA) was added to these sections.

Techniques:

Journal: BMC Women's Health

Article Title: AlphaVBeta3 Integrin expression within uterine endometrium in unexplained infertility: a prospective cohort study

doi: 10.1186/s12905-017-0438-3

Figure Lengend Snippet: Immunostained slide with anti- vβ3 integrin, DAB chromogen, magnification x 200 (High Power), score 3 in A case of fertility group. Reactivity could be seen Mainly in Luminal Epithelium (red arrow) rather than the Glandular Epithelium (red star)

Article Snippet: After blocking with non-immune 4% goat serum incubated with tissues for 30 min, the primary antibody using monoclonal mouse IgG antibodies directed against human αvβ3 integrin (Human integrin alphaV beta3 MAb (clone 23C6)®, MAB3050, R&D systems, Minneapolis, USA) was added to these sections.

Techniques:

Journal: BMC Women's Health

Article Title: AlphaVBeta3 Integrin expression within uterine endometrium in unexplained infertility: a prospective cohort study

doi: 10.1186/s12905-017-0438-3

Figure Lengend Snippet: Comparison of endometrial thickness, subendometrial Doppler resistance index, and ανβ3-integrin score in both study groups

Article Snippet: After blocking with non-immune 4% goat serum incubated with tissues for 30 min, the primary antibody using monoclonal mouse IgG antibodies directed against human αvβ3 integrin (Human integrin alphaV beta3 MAb (clone 23C6)®, MAB3050, R&D systems, Minneapolis, USA) was added to these sections.

Techniques: Comparison, Control

Journal: BMC Women's Health

Article Title: AlphaVBeta3 Integrin expression within uterine endometrium in unexplained infertility: a prospective cohort study

doi: 10.1186/s12905-017-0438-3

Figure Lengend Snippet: Receiver-operating characteristic (ROC) curve for the discrimination between patients with unexplained infertility and normal controls using a : Endometrial thickness, b : Subendometril RI, c : avβ3 integrin

Article Snippet: After blocking with non-immune 4% goat serum incubated with tissues for 30 min, the primary antibody using monoclonal mouse IgG antibodies directed against human αvβ3 integrin (Human integrin alphaV beta3 MAb (clone 23C6)®, MAB3050, R&D systems, Minneapolis, USA) was added to these sections.

Techniques:

Journal: BMC Women's Health

Article Title: AlphaVBeta3 Integrin expression within uterine endometrium in unexplained infertility: a prospective cohort study

doi: 10.1186/s12905-017-0438-3

Figure Lengend Snippet: Receiver-operating characteristic (ROC) curve analysis for the discrimination between patients with unexplained infertility and control group

Article Snippet: After blocking with non-immune 4% goat serum incubated with tissues for 30 min, the primary antibody using monoclonal mouse IgG antibodies directed against human αvβ3 integrin (Human integrin alphaV beta3 MAb (clone 23C6)®, MAB3050, R&D systems, Minneapolis, USA) was added to these sections.

Techniques: Control

Journal: BMC Women's Health

Article Title: AlphaVBeta3 Integrin expression within uterine endometrium in unexplained infertility: a prospective cohort study

doi: 10.1186/s12905-017-0438-3

Figure Lengend Snippet: Summary of studies looked at endometrial αVβ3 integrin in luteal phase of infertile women

Article Snippet: After blocking with non-immune 4% goat serum incubated with tissues for 30 min, the primary antibody using monoclonal mouse IgG antibodies directed against human αvβ3 integrin (Human integrin alphaV beta3 MAb (clone 23C6)®, MAB3050, R&D systems, Minneapolis, USA) was added to these sections.

Techniques: Control, Expressing

Journal: RSC Advances

Article Title: RGD-modified dihydrolipoamide dehydrogenase conjugated to titanium dioxide nanoparticles – switchable integrin-targeted photodynamic treatment of melanoma cells †

doi: 10.1039/c7ra13777j

Figure Lengend Snippet: Cytotoxic effect of the nanobiocomplex (A-NPs–DLDH RGD ) in the presence or absence of UVA illumination. B16F10 or HEK293 cells were treated with TiO 2 , TiO 2 –DLDH or TiO 2 –DLDH RGD and assessed after an overnight by confocal microscopy (Leica SP5) under (A) dark conditions or (B) UVA illumination (1 h, 365 nm). Cells treated in medium lacking TiO 2 or the DLDH forms, served as controls. The cell nuclei were stained red (Draq5). Experiments were repeated twice in triplicates. Next the same experiment was analyzed for cell counts under (C) dark conditions or (D) UVA illumination. Experiments were repeated 3 times in triplicates. Significance (* p < 0.05, ** p < 0.005, *** p < 0.001) from control cells after uva illumination. Significance between the experiments groups (±RGD, ±αvβ3) are indicated by an horizontal line.

Article Snippet: The cells were harvested in RPMI 1640 and labeled with 10 μg ml −1 FITC-αvβ3 antibody (clone LM609, Merck Millipore, Darmstadt, Germany).

Techniques: Confocal Microscopy, Staining, Control

Journal: Cell Adhesion & Migration

Article Title: Selective binding and lateral clustering of α 5 β 1 and α v β 3 integrins: Unraveling the spatial requirements for cell spreading and focal adhesion assembly

doi: 10.1080/19336918.2016.1163453

Figure Lengend Snippet: Cell spreading kinetics on nanopatterned surfaces functionalized with integrin selective ligands. (A) Progression of projected cell area during spreading on nanopatterned surfaces with interparticle distances of 30, 60, or 90 nm, and functionalized with α5β1 (white) and αvβ3 (black) integrin selective ligands. (B) Maximum projected cell area on the different surfaces. Error bars indicate SEM of 3 independent repeats.

Article Snippet: Cells were then washed with warm PBS, and fixed in 3.7% paraformaldehyde in PBS for 20 min. Post-fixation, cells were permeabilized with 0.1% TritonX-100 in PBS for 5 min, blocked with 1% bovine serum albumin (BSA) in PBS, and incubated for 1 hr with the following antibodies: mouse anti-human αvβ3 integrin (Millipore, MAB1976), rat anti-human α5-integrin (MABII, kindly provided by K. Yamada), mouse anti- human vinculin (Sigma, V9131), and rabbit anti-human zyxin (Synaptic Systems, 307011).

Techniques:

Journal: Cell Adhesion & Migration

Article Title: Selective binding and lateral clustering of α 5 β 1 and α v β 3 integrins: Unraveling the spatial requirements for cell spreading and focal adhesion assembly

doi: 10.1080/19336918.2016.1163453

Figure Lengend Snippet: Focal adhesions in cells adhering to nanopatterned surfaces functionalized with integrin α5β1 and αvβ3 integrin selective ligands. (A) Indirect immunofluorescence staining of vinculin (green), phosphorylated paxillin (red), and actin (blue) in U2OS cells. Insets are a magnification of separate stainings for vinculin and phosphorylated paxillin, in the cell region delineated by the white box. Cells adhering for 4 hr to α5β1 (first row) and αvβ3 integrin selective ligands (second row) at spacings of 30 nm (left), 60 nm (middle), and 90 nm (right) were imaged by wide-field microscopy. (B) Analysis of vinculin cluster size; and (C) Analysis of phosphorylated paxillin (PY118) cluster size in U2OS cells. Box plots indicate cluster area values between 25% and 75%, and whiskers between 10% and 90% of the data range. The line in the box plot indicates the median value. p * < 0.001.

Article Snippet: Cells were then washed with warm PBS, and fixed in 3.7% paraformaldehyde in PBS for 20 min. Post-fixation, cells were permeabilized with 0.1% TritonX-100 in PBS for 5 min, blocked with 1% bovine serum albumin (BSA) in PBS, and incubated for 1 hr with the following antibodies: mouse anti-human αvβ3 integrin (Millipore, MAB1976), rat anti-human α5-integrin (MABII, kindly provided by K. Yamada), mouse anti- human vinculin (Sigma, V9131), and rabbit anti-human zyxin (Synaptic Systems, 307011).

Techniques: Immunofluorescence, Staining, Microscopy

Journal: Cell Adhesion & Migration

Article Title: Selective binding and lateral clustering of α 5 β 1 and α v β 3 integrins: Unraveling the spatial requirements for cell spreading and focal adhesion assembly

doi: 10.1080/19336918.2016.1163453

Figure Lengend Snippet: α5 and αvβ3 clusters in U2OS cells adhering to nanopatterned surfaces functionalized with α5β1 and αvβ3 integrin selective ligands. (A) Cells adhering to surfaces with 30 nm interparticle spacing; and (B) Cells adhering to surfaces with 60 nm particle spacing. Upper row: Cells adhering to α5β1 integrin selective ligands. Lower row: Cells adhering to αvβ3 integrin selective ligands. Left: Staining for α5 clusters. Middle: Staining for αvβ3 clusters. Right: Lookup table displaying the colocalization of α5 and αvβ3 integrin clusters (pixel with positive signals for both integrins are shown in yellow).

Article Snippet: Cells were then washed with warm PBS, and fixed in 3.7% paraformaldehyde in PBS for 20 min. Post-fixation, cells were permeabilized with 0.1% TritonX-100 in PBS for 5 min, blocked with 1% bovine serum albumin (BSA) in PBS, and incubated for 1 hr with the following antibodies: mouse anti-human αvβ3 integrin (Millipore, MAB1976), rat anti-human α5-integrin (MABII, kindly provided by K. Yamada), mouse anti- human vinculin (Sigma, V9131), and rabbit anti-human zyxin (Synaptic Systems, 307011).

Techniques: Staining

Journal: Cell Adhesion & Migration

Article Title: Selective binding and lateral clustering of α 5 β 1 and α v β 3 integrins: Unraveling the spatial requirements for cell spreading and focal adhesion assembly

doi: 10.1080/19336918.2016.1163453

Figure Lengend Snippet: α5β1 and αvβ3 integrin blocking. (A) Phase contrast micrographs of U2OS cells incubated with α5β1 and αvβ3 integrin selective ligands, and seeded on nanopatterned surfaces functionalized with these ligands. Upper row: Cells adhering to α5β1integrin selective ligands. Lower row: Cells adhering to αvβ3 integrin selective ligands. Left: No integrin blocking. Middle: α5β1 integrin blocking. Right: αvβ3 integrin blocking. (B) Indirect immunofluorescence staining of α5 (green) and αvβ3 clusters (red) in U2OS cells pre-incubated with the integrin selective ligands. Cells were seen to adhere to nanopatterned surfaces functionalized with α5β1 (upper row) and αvβ3 integrin selective ligands (lower row).

Article Snippet: Cells were then washed with warm PBS, and fixed in 3.7% paraformaldehyde in PBS for 20 min. Post-fixation, cells were permeabilized with 0.1% TritonX-100 in PBS for 5 min, blocked with 1% bovine serum albumin (BSA) in PBS, and incubated for 1 hr with the following antibodies: mouse anti-human αvβ3 integrin (Millipore, MAB1976), rat anti-human α5-integrin (MABII, kindly provided by K. Yamada), mouse anti- human vinculin (Sigma, V9131), and rabbit anti-human zyxin (Synaptic Systems, 307011).

Techniques: Blocking Assay, Incubation, Immunofluorescence, Staining

Journal: PLoS ONE

Article Title: High Glucose Alters Retinal Astrocytes Phenotype through Increased Production of Inflammatory Cytokines and Oxidative Stress

doi: 10.1371/journal.pone.0103148

Figure Lengend Snippet: Integrins α5β1 (A) and αvβ3 (B) levels were determined by FACS analysis. Shaded areas show staining in the absence of primary antibody. Please note the dramatic decrease (2-fold) in mean florescence intensity of α5β1 integrin in AC under high glucose conditions compared to normal glucose and osmolarity control conditions (P<0.05, n = 3). This was mainly attributed with a nearly 2-fold decrease in the fluorescence intensity of α5 integrin, while the fluorescence intensity of β1 integrin did not change under various glucose conditions (not shown). The mean fluorescence intensity of αvβ3 was increased (1.5-fold) under high glucose or osmolarity control conditions compared to normal glucose conditions (P<0.05 n = 3), consistent with enhanced adhesion of AC to fibronectin and vitronectin under high glucose conditions.

Article Snippet: The anti-cleaved caspase 3 antibody (Cell signaling, Danvers, MA), anti-GFAP (Dako, Carpentaria, CA), anti-α5β1 integrin (MAB1976Z), or anti-αvβ3 integrin (MAB1999) (Millipore, Temecula, CA) was prepared in TBS with 1% BSA at 2 μg/ml.

Techniques: Staining, Control, Fluorescence

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Self-Triggered Apoptosis Enzyme Prodrug Therapy (STAEPT): Enhancing Targeted Therapies via Recurrent Bystander Killing Effect by Exploiting Caspase-Cleavable Linker.

doi: 10.1002/advs.201800368

Figure Lengend Snippet: Figure 2. Enhanced cellular uptake and cytotoxic activity of RGDEVD-DOX in integrin αvβ3 expressing cells. a) Representative confocal images (left) and quantitative analysis (right) of HDMEC and U-87 MG cells exposed to fluorescent-labeled RDEVD and RGDEVD peptides. b) Representative confocal images (left) and quantitative analysis (right) of scrambled control and integrin αv (ITGAV) siRNA-transfected HDMECs and U-87 MG cells exposed to fluorescent-labeled RGDEVD peptide. Green and blue indicate the fluorescent-labeled peptides and cell nuclei, respectively. Scale bar, 50 µm. c) Flow cytometry analysis of isotype control (top), scrambled control-transfected (lower left), and ITGAV siRNA-transfected (lower right) U87 MG cells incubated with fluorescent-labeled RGDEVD and stained with an antibody against integrin αvβ3. d) Representative confocal images (left) and quantitative analysis (right) of U-87 MG and HT-29 cells treated with RDEVD-DOX and RGDEVD-DOX. Red and blue indicate the intrinsic red fluorescence of doxorubicin and cell nuclei, respectively. Scale bar, 50 µm. e) Concentration-dependent cytotoxicity of RDEVD-DOX and RGDEVD-DOX on U-87 MG (left) and HT-29 (right) determined by MTT assay (n = 4). f) Doxorubicin release from RGDEVD-DOX when incubated in PBS (pH 7.4) containing (or not containing) carboxylesterase (n = 3). Data are mean ± s.d. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: The cells were incubated with Alexa Fluor 488-conjugated integrin αvβ3 antibody (1:100; R&D Systems, Minneapolis, MN; Cat. No. FAB3050G) for an hour at 4 °C, washed, and suspended in PBS containing 0.5% BSA.

Techniques: Activity Assay, Expressing, Labeling, Control, Transfection, Flow Cytometry, Incubation, Staining, Fluorescence, Concentration Assay, MTT Assay