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| Description | Movie of an NIH 3T3 mouse fibroblast that has been starved for growth factors and then stimulated by platelet-derived growth factor (PDGF). The cell is expressing a reporter for PIP3 (phosphatidylinositol(3, 4, 5)trisphosphate), composed of a PIP3 binding domain (the pleckstrin homology domain of Bruton's Tyrosine Kinase) fused to enhanced green fluorescent protein (EGFP). In the starved cell the reporter is partially nuclear (likely due to a weak nuclear localization sequence within EGFP). Beginning at the third frame, in response to 50ng/ml PDGF, the reporter rapidly reveals the production of the lipid second messenger PIP3 at the plasma membrane. |
| Title | PIP3 Produced in Response to Stimulation by a Growth Factor |
| Author(s) | Seth J Field |
| Materials & Methods | NIH 3T3 mouse fibroblasts were transfected with expression vectors for BTK-PH-EGFP (reporter for PIP3) and dsRed (used to normalize for cell volume). Cells were serum-starved overnight, and then imaged live at 37C. Images of BTK-PH-EGFP and dsRed fluorescence were captured every 40 seconds. At the third frame PDGF was added to the media to a final concentration of 50ng/ml, while image capturing continued. Ultimately, ratio images of BTK-PH-EGFP/dsRed were calculated, representing a measure of BTK-PH-EGFP per unit volume (although un-normalized images look roughly similar to the normalized images). The scale bar indicates 10 microns. The time counter indicates time in seconds, time zero corresponds to the time of addition of PDGF. |
| Citation | Field S. PIP3 produced in response to stimulation by a growth factor. ASCB Image & Video Library. August 2007:VID-28. Available at: http://cellimages.ascb.org |
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| Publication Date | Published in August 2007. |
| Contributors | Work conducted at University of California, San Diego, La Jolla, CA. |
| Resource Type | Video |
| Digital Format | quicktime; Digitization process: Recompressed with H.264 codec, automatic keyframes selected and data rate restricted to 2000 kbs (optimized for download) in Quicktime Pro. New file size: 27.5 kb; Duration: 00:00:07.00; Dimensions: 175 x 218; FPS: 3; Data rate: 3.94 kb; Codec: H.264; Software required: Apple Quicktime 7 player: http://www.apple.com/quicktime/download/win.html. |
| Original Date | Original movie created on January 16, 2001. |
| Original Format | Original resource: a series of TIFF images that were combined into a Quicktime movie. Received from authors: quicktime; File size: 120 kb; Duration: 00:00:07.00; Dimensions: 175 x 218 pixels; FPS: 3; Data rate: 131.51 kb/sec; Codec: Apple MPEG4. |
| License Details | Terms for non-commercial use: http://cellimages.ascb.org/cdm4/terms.php; for commercial use contact cellimages@ascb.org. |
| Publisher | The American Society for Cell Biology |
