Uev1A, a ubiquitin conjugating enzyme variant, inhibits stress-induced apoptosis through NF-kappaB activation.

We have previously shown that UEV1 is up-regulated in all tumor cell lines examined and when SV40-transformed human embryonic kidney cells undergo immortalization; however, it is unclear whether and how UEV1 plays a critical role in this process. UEV1A encodes a ubiquitin conjugating enzyme variant, which is required for Ubc13 (ubiquitin conjugating enzyme) catalyzed poly-ubiquitination of target proteins through Lys63-linked chains. One of the target proteins is NEMO/IKKgamma (nuclear factor-kappaB essential modulator/inhibitor of kappaB protein kinase), a regulatory subunit of IkappaB kinase in the NF-kappaB signaling pathway.

A simple, informative, and quantitative flow cytometric method for assessing apoptosis in cultured cells.

Described herein is a method for assessing apoptosis in tissue culture cells that is facile, highly informative, readily quantifiable, and generally applicable. As in conventional DNA-based flow cytometric analysis, the approach utilizes fixed, propidium iodide-stained cells. However, this particular application employs correlated two-parameter analyses of log(10)DNA fluorescence signals versus log(10) side-scatter (SS) signals of cells undergoing apoptosis.