Under copyright Constraint(s) on Use: This work is protected by the U.S. Copyright Law (Title 17, U.S.C.). Use of this work beyond that allowed by "fair use" or any license applied to this work requires written permission of the copyright holder(s). Responsibility for obtaining permissions and any use and distribution of this work rests exclusively with the user and not the UC San Diego Library. Inquiries can be made to the UC San Diego Library program having custody of the work. Use: This work is available from the UC San Diego Library. This digital copy of the work is intended to support research, teaching, and private study.
Rights Holder and Contact
UC Regents
Description
TNF-alpha stimulated p110alpha or control siRNA transfected HUVECs previously labelled with celltracker orange dye were grown on collagen matrices in transwell chambers. Celltracker green-labelled THP-1 cells were added to HUVECs, and TEM allowed to proceed toward an MCP-1 gradient in the lower chamber, for either 10 or 60 minutes before fixation. Fixed samples were stained with AlexaFluor 633-conjugated phalloidin to visualize F-actin (grey) and z-stacks collected. Three dimensional reconstructions were produced using Volocity software. A small gamma correction was applied to the red channel in order to visualise weak cell tracker staining at the edges of cell, which are difficult to distinguish upon 3D rendering. Transwells were fixed by immersion in 3.7% (wt/vol) paraformaldehyde before being processed for immunofluorescence microscopy. Images were acquired at room temperature using a confocal microscope (LSM 510 META; Carl Zeiss, Inc.) with three single photomultiplier tube confocal detectors mounted on an inverted microscope (AxioObserver Z1; Carl Zeiss, Inc.) at a magnification of 40 with an oil immersion objective (EC Plan-Neofluar 40× NA 1.30 oil differential interference contrast M27). z sections were acquired at 200-nm intervals by confocal microscopy. Corresponds to Figure 5 from J Cell Biol. 2010 Mar 22;188(6):863-76. Research Data Curation Program, UC San Diego, La Jolla, 92093-0175 (https://lib.ucsd.edu/rdcp) Cain, Robert J.; Vanhaesebroeck, Bart; Ridley, Anne J. (2021). CIL:30547, Homo sapiens, blood vessel endothelial cell, monocytic leukemia cell. In Cell Image Library. UC San Diego Library Digital Collections. Dataset. https://doi.org/10.6075/J0GM85X6
If you're wondering about permissions and what you can do with this item, a good starting point is the "rights information" on this page. See our terms of use for more tips.
Share your story
Has Calisphere helped you advance your research, complete a project, or find something meaningful? We'd love to hear about it; please send us a message.
