Journal: Molecular Cancer Research

Article Title: β8 Integrin Mediates Pancreatic Cancer Cell Radiochemoresistance

doi: 10.1158/1541-7786.mcr-18-1352

Figure Lengend Snippet: Figure 5. b8 integrin interactome and subcellular colocalization. A, Sequential immunoprecipitation-mass spectrometry–based analysis of b8 integrin interactome was performed in unirradiated and 6-Gy X-rays irradiated Patu8902 cells. Time point of analysis after irradiation was 2 hours. B, Comparative changes in the b8 integrin interactome between control and irradiation upon categorization into different molecular functions using the PANTHER classification system. C, Immunofluorescence costaining of b8 integrin with GM130 (Golgi), mitochondria, aV integrin, APPL2, and Caveolin 1. Scale bars, 10 mm. D, Pearson correlation analysis of C. Data show mean SD (n ¼ 3).

Article Snippet: The antibodies against b1 integrin (WB 1:1,000, Abcam, ab179471), b8 integrin (IF 1:200, WB 1:1,000; Abcam, ab80673, Rb, recognizes aa 614-663 (this antibody was generally used in this study); Abnova, H00003696-M01, Mo, recognizes aa 392–503 (this antibody was used only where specifically indicated), GM130 (IF 1:100, WB 1:1,000, BD, 610822), MEK1/2 (WB 1:1,000, Cell Signaling Technology, 4694s), gH2AX (WB 1:1,000, Cell Signaling Technology, 9718s), aV integrin (IF 1:250, Novus, NB100-2618), APPL2 (IF 1:100, Sigma-Aldrich, SAB1400605), Caveolin 1 (IF 1:100, BD, 610407), LC3B (IF 1:500, Sigma-Aldrich, SAB4200361), b-actin (WB 1:10,000, Sigma-Aldrich, A5441), HRP-conjugated secondary antibody (GE, Rb NXA931, Mo NXA931), Alexa Fluor 488– or Alexa Fluor 594–conjugated secondary antibodies (1:500, Life Technologies) were purchased and used as indicated.

Techniques: Immunoprecipitation, Mass Spectrometry, Irradiation, Control

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

Journal: Biochimie

Article Title: Inhibition of platelets and tumor cell adhesion by the disintegrin domain of human ADAM9 to collagen I under dynamic flow conditions.

doi: 10.1016/j.biochi.2009.05.012

Figure Lengend Snippet: Fig. 5. ADAM9D binds to MDA-MB-231 tumor breast cells by b1, a3, aVb3, aVb5 and a2 integrins. Ninety six well plates were coated with ADAM9D (10 mg/ml) overnight at 4 C. After blocking with 1% BSA, the CMFDA-labeled MDA-MB-231 cells (1 105 cells/ well) previously incubated (30 min, 37 C) or not (control bar) with different anti- bodies (20 mg/well) were added to each well. The plates were incubated at 37 C for 30 min, washed, lysed and read as in Fig. 2. Results are expressed as mean SEM of three independent experiments. The results were normalized by the ADAM9D (control) values which were considered as 100% adhesion The p value was determined using the using Dunnett’s test comparing antibodies bars with the control bar (*p < 0.05; **p < 0.01; ***p < 0.001).

Article Snippet: For competition assays the monoclonal antibodies directed against human a2 (MAB1233), a6 (MAB13501), b1 (MAB17781) and aVb5 (MAB2528) integrins were obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Blocking Assay, Labeling, Incubation, Control