Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2003 Aug 18;162(4):587–597. doi: 10.1083/jcb.200305149

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

Copyright © 2003, The Rockefeller University Press

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

PMC Copyright notice

Figure 7. — Expression of bbc3/PUMA may be sufficient and required for ER stress–induced apoptosis in human cells. (A) Transient transfection of SH-SY5Y neuroblastoma cells with bbc3/PUMA leads to induction of apoptosis. Cells were cotransfected with expression plasmids encoding enhanced GFP (pEGFP) and either a hemagglutinin-tagged (HA) version of bbc3/PUMA (pHA-PUMA) or empty vector (pcDNA3.1). 12 h after transfection, cells were fixed and stained with Hoechst 33258. Arrowheads point to transfected cells with the apoptotic phenotype. Similar results were obtained in a separate experiment. (B) Quantitative FACS^® analysis of PI uptake of EGFP-positive SH-SY5Y cells transiently cotransfected with expression vectors encoding hemagglutinin-tagged (HA) versions of Bbc3/PUMA, Bbc3/PUMA-ΔBH3, or empty vector (pcDNA3.1). Data were normalized to EGFP-only transfected controls and represent 10,000 events each. For evaluation of comparable transfection rates, whole-cell extracts of cells harvested 16 h after transfection were analyzed by Western blotting. Blots were probed with an anti-hemagglutinin or anti- α-tubulin antibody as loading control. (C) Protein levels of BiP, Bbc3/PUMA, and α-tubulin in human HCT116 control and Bbc3/PUMA-deficient cells treated with 1 μM thapsigargin or vehicle for the indicated time points. (D) FACS^® analysis of sub-G1 cells of human HCT116 control and Bbc3/PUMA-deficient cells exposed to 1 μM thapsigargin (TH) or vehicle for 36 h. (E) Quantitative FACS^® analysis of thapsigargin- and STS (3 μM)-induced apoptosis. Data from 10,000 events each are means ± SEM from n = 3 separate experiments per time point.