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Image Search Results
Journal: Oncotarget
Article Title: Discoidin domain receptor 1 promotes Th17 cell migration by activating the RhoA/ROCK/MAPK/ERK signaling pathway.
doi: 10.18632/oncotarget.10455
Figure Lengend Snippet: Figure 2: DDR1 promotes human Th17 motility in 3D collagen. A. DDR1 expression is reduced by specific DDR1 siRNA (HSS1878780). Polarized Th17 cells were transfected with control or with DDR1 siRNAs and DDR1 expression was assessed by western blot using the anti-DDR1 antibody (C-20). The blot was stripped and reprobed with anti-β-actin antibody to ensure equal loading (top panel). After transfection, the cells were activated with PMA+ionomycin in the presence of brefeldin A to identify IL-17-producing cells, stained with anti-DDR1 and anti-IL-17 antibodies and analyzed by flow cytometry (lower panel). Immunoblot and FACS plots are representative of five independent experiments performed with polarized Th17 cells derived from five different blood donors. B. DDR1 siRNA inhibits Th17 motility in 3D collagen. After transfection, the cells were labelled with calcein AM and embedded in collagen gels. Cell migration was evaluated by live cell confocal microscopy and quantified by computer-assisted cell tracking as described in the “Materials and Methods” section. Representative cell migration tracks over 30 min are presented as x-y projections (distance, in μm) (left panel). The histogram (right panel) represents the mean velocity of 100 cells presented as μm/min. C. DDR1:Fc inhibits Th17 motility in 3D collagen. The cells were embedded in collagen gels containing control human IgG (Fc fragment) or human DDR1:Fc recombinant proteins and cell motility was determined as above. Results (B and C right panels) are mean values ± SD of five independent experiments performed with polarized Th17 cells derived from five different blood donors. *p < 0.05. D. DDR1:Fc inhibits the migratory shape of polarized Th17 cells. Representative photography images from five different experiments of polarized Th17 cells migrating in collagen gels containing either control IgG or DDR1:Fc (400X magnification).
Article Snippet: Non-conjugated rabbit anti-mouse and human DDR1 (clone C-20), non-conjugated rabbit anti-human DDR2 (clone H108) antibodies and the anti-phospho-ERK1/2 (clone E-4), anti-ERK2 (clone C-14), and
Techniques: Expressing, Transfection, Control, Western Blot, Staining, Flow Cytometry, Derivative Assay, Migration, Confocal Microscopy, Cell Tracking Assay, Recombinant