{"CIL_CCDB":{"Status":{"Is_public":true,"Deleted":false,"Publish_time":1306123200},"Data_type":{"Still_image":true,"Z_stack":false,"Video":false,"Time_series":false},"CIL":{"Image_files":[{"File_type":"Zip","File_path":"26536.zip","Size":792100,"Mime_type":"application\/zip"},{"File_type":"OME_tif","File_path":"26536.tif","Size":800000,"Mime_type":"image\/tif"},{"File_type":"Jpeg","File_path":"26536.jpg","Size":49279,"Mime_type":"image\/jpeg; charset=utf-8"}],"CORE":{"ATTRIBUTION":{"URLs":[{"Href":"http:\/\/jcb-dataviewer.rupress.org\/jcb\/browse\/3112\/"}],"PUBLISHED":["J. Cell Biol. 2010. Vol. 190(4):663\u2013674"],"Contributors":["Chan-Soo Lee","Chang-Ki Choi","Eun-Young Shin","Martin Alexander Schwartz","Eung-Gook Kim"],"PUBMED":["PMID:20713598"]},"BIOLOGICALPROCESS":{"onto_name":"regulation of cell migration","onto_id":"GO:0030334"},"CELLLINE":{"onto_name":"NIH\/3T3","onto_id":"MCC:0000362"},"PARAMETERIMAGED":{"onto_name":"fluorescence emission","onto_id":"FBbi:00000316"},"IMAGEDESCRIPTION":{"free_text":"To study the molecular mechanism by which nonmuscle myosin II (MII) regulates protrusion and adhesion dynamics in migrating cells, NIH3T3 cells were transfected with myc-tagged phosphomimetic constitutively active MLC mutants (MLCee, green).  After 20' exposure to PDGF, which induces myosin-inactivation, stress fiber disassembly, and stimulates fibroblast motility, the cells were fixed and co-stained for betaPIX (a Rac1\/Cdc42-specific GEF highly implicated in cell motility, red) and filamentous actin (blue). These findings help elucidate a functional link between MII and Rac1\/Cdc42 GTPases, which may regulate protrusion\/adhesion dynamics in migrating cells.  This image is original data file Fig. 7C \"PDGF-induced dissociation of the MII\u2013βPIX complex,\" in J. Cell Biol. 2010. Vol. 190(4):663\u2013674"},"ITEMTYPE":{"onto_name":"recorded image","onto_id":"FBbi:00000265"},"CELLULARCOMPONENT":[{"onto_name":"lamellipodium","onto_id":"GO:0030027"},{"onto_name":"myosin II complex","onto_id":"GO:0016460"},{"onto_name":"actin filament","onto_id":"GO:0005884"}],"RELATIONTOINTACTCELL":{"onto_name":"dispersed cells in vitro","onto_id":"FBbi:00000611"},"SOURCEOFCONTRAST":[{"onto_name":"distribution of epitope","onto_id":"FBbi:00000592"},{"onto_name":"distribution of a specific protein","onto_id":"FBbi:00000597"}],"GROUP_ID":"8283","TECHNICALDETAILS":{"free_text":"Cells were cultured in DME (Invitrogen) supplemented with 10% fetal bovine serum and 100 U\/ml penicillin\/streptomycin (Invitrogen) at 37°C in a humidified 5% CO2 incubator. For transfections, cells in 60-mm-diameter dishes or on fibronectin-coated coverslips were incu- bated with a mixture of DNA and LipofectAMINE 2000 (Invitrogen) according to the manufacturer\u2019s instructions. The cDNA for MLC was subcloned into pCMV-myc (Takara Bio Inc.). Mutant constructs of MLC were generated using a QuikChange site-directed mutagenesis kit (Agilent Technologies) according to the manufacturer\u2019s protocol. In this myc-tagged MLC mutant, Thr18\/Ser19 were replaced by glutamic acids, and expressed in NIH3T3 cells.\nIn some experimental conditions, 16 h after replating onto fibronectin-coated coverslips, cells were treated with 50 ng\/ml PDGF for 20 min.  Cells were fixed 24\u201348 h after transfection using 3.7% paraformaldehyde in PBS for 15 min, permeabilized using 0.2% Triton X-100 in PBS for 2 min, blocked with 2% BSA in PBS, and co-stained for MLC (green), TRIO (red), and actin (blue). Images were captured by Zeiss LSM 710 confocal microscope with Plan-Apochromat 63X objective."},"DATAQUALIFICATION":{"free_text":"RAW"},"TERMSANDCONDITIONS":{"free_text":"attribution_nc_sa"},"IMAGINGMODE":{"onto_name":"confocal microscopy","onto_id":"FBbi:00000251"},"DIMENSION":[{"Space":{"Image_size":512,"axis":"X"}},{"Space":{"Image_size":512,"axis":"Y"}}],"NCBIORGANISMALCLASSIFICATION":{"onto_name":"Mus musculus","onto_id":"NCBITaxon:10090"},"PREPARATION":[{"onto_name":"formaldehyde fixed tissue","onto_id":"FBbi:00000010"},{"onto_name":"detergent permeabilized","onto_id":"FBbi:00000262"}],"VISUALIZATIONMETHODS":[{"onto_name":"Alexa Fluor 488","onto_id":"FBbi:00000440"},{"onto_name":"Alexa Fluor 546","onto_id":"FBbi:00000441"},{"onto_name":"phalloidin","onto_id":"FBbi:00000100"}],"CELLTYPE":{"onto_name":"fibroblast","onto_id":"CL:0000057"}}},"Citation":{"Title":"Chan-Soo Lee, Chang-Ki Choi, Eun-Young Shin, Martin Alexander Schwartz, Eung-Gook Kim (2011) CIL:26536, Mus musculus, fibroblast. CIL. Dataset","ARK":"ark:\/b7295\/w9cil26536","DOI":"doi:10.7295\/W9CIL26536"}}}