antibodies against integrin α 5 β v  (R&D Systems)

Journal: Science Advances

Article Title: Integrin inhibition facilitates fibrocartilaginous transformation in connective tissue in osteoarthritis

doi: 10.1126/sciadv.ady4112

Figure Lengend Snippet: ( A ) Dot plot shows notably increased ECM-related signaling pathways of FB4 ( COL5A1 + ) in ADD TMJ-OA RCT compared to normal RCT. ( B ) GO analysis shows potentially up-regulated gene functions of FB4 ( COL5A1 + ) in ADD TMJ-OA RCT compared to normal RCT. Top 15 GO terms of the enrichment factors are showed. MHC, major histocompatibility complex; ER, endoplasmic reticulum. ( C ) Dot plot shows the expression of integrin-related genes in FB4 ( COL5A1 + ). The red (blue) dots are the up-regulated (down-regulated) genes in ADD TMJ-OA RCT compared to normal RCT. Horizontal and vertical coordinates indicate the specificity of the α and β integrin subunit genes on FB4 ( COL5A1 + ) cells. ( D ) The chord diagram shows the ligand-receptor pairs of periostin signaling (left). Proportions of periostin signaling interaction strength are compared between normal RCT and ADD TMJ-OA RCT (right). ( E ) Left plot shows coexpression of ITGAV and ITGB5 in FB4 ( COL5A1 + ) cells on the 10x Visium capture slides. Proportions of the coexpression of ITGAV and ITGB5 in FB4 ( COL5A1 + ) cells are compared between normal RCT and ADD TMJ-OA RCT. ( F ) Immunofluorescent staining for integrin α V β 5 in human TMJ disk tissues. Red: integrin α V β 5 ; blue: DAPI. White dotted lines: boundary of the disk tissues. N = 3 independent samples from different donors. Scale bars, 500 μm (left) and 50 μm (right). ( G ) Semiquantification of the percentage of integrin α V β 5 + area. Data are presented as mean ± SD. N = 3 independent animals. The two-tailed t test is used for data analysis. ** P < 0.01.

Article Snippet: Primary antibodies, which included antibodies against integrin α 5 β V (MAB2528, AB_2280706, R&D Systems, USA) diluted to 1:100 in blocking solution, were incubated at 4°C overnight.

Techniques: Protein-Protein interactions, Immunopeptidomics, Expressing, Staining, Two Tailed Test

Journal: Journal of Oncology

Article Title: Anoikis Resistance as a Further Trait of Acidic-Adapted Melanoma Cells

doi: 10.1155/2019/8340926

Figure Lengend Snippet: An anoikis resistant-like phenotype in acidic melanoma cells. (a) Representative images of western blot for N-cad, EGFR, pAKT/AKT, pERK/ERK, Tie1, IKB, and β -actin of A375M6 melanoma cells exposed to standard medium (pH 7.4), to an acidified medium for 24 hours (transient exposure, pH 6.7) or to a reduced pH medium, for approximately three months (chronic exposure, pH 6.7c), and (right) densitometry graph of protein expression. (b) Representative images (left) of invasiveness of melanoma cells grown in different pH conditions in the presence or absence of 25 μ M Ilomastat (a metalloproteinases inhibitor) and quantitative analysis (right) of the number of cells that migrated through Matrigel. (c) Representative images (left) of flow cytometric analysis of α v β 3 and of α v β 5 integrin expression of A375M6 melanoma cells grown in different pH conditions and (right) quantitative analysis of integrin expression as percentage of increment in Mean Fluorescent Intensity. Each experiment was conducted in triplicate and data are expressed as mean ± SEM of at least three independent experiments. ∗ p<0.05.

Article Snippet: Mouse anti-human integrin receptor α v β 3 (clone LM609, Millipore) and mouse anti-human integrin receptor α v β 5 (sc13588, Sigma), mouse monoclonal AF6-88.5.3 specific for class I H-2Kb antigen (provided by Dr. S. Gattoni-Celli, Medical University of South Carolina, USA) (Calorini 1999) or mouse monoclonal HLA-ABC antigen clone W6/32 (DAKO), were used.

Techniques: Western Blot, Expressing