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

. 2007 Oct 8;179(1):23–31. doi: 10.1083/jcb.200706097

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

Copyright © 2007, 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 1. — p66^Shc confers anoikis. (A) 293 cells transiently transfected with pCINF-p52 or stably transfected with an empty vector, pCINF-p66 wt, pCINF-p66 (S36A), or p66 (S36E) were left adherent or suspended for 16 h in low-attachment plates. Nuclear DNA fragmentation was assessed and demonstrated increased cell death with detachment in cells expressing p66 (wt) or p66 (SA). (inset) Immunoblot for Shc showing p66^Shc, p52^Shc, and p46^Shc expression. *, P < 0.001 compared with attached control; †, P < 0.001 compared with p66 (wt) floating. (B) Stable transfectants of empty vector (293-vector) or p66^Shc (293-p66) were fractionated into mitochondrial or cytosolic content after 16 h of adherent or suspended culture and immunoblotted with the indicated antibodies. Bim was strongly induced and remained in the mitochondrial fraction with detachment. (C) 293 cells expressing either p66 wt or p66 (S36A) were immunoblotted after 16 h of suspension or adherence for phospho-S36 or total p66^Shc. (D) Stable p66^Shc cells were transiently transfected with either vector or the isolated Shc CH2 domain, and DNA fragmentation was measured after 16 h of adherence or suspension. (E) HUVEC were transfected with either control siRNA or siRNA against the CH2 domain of Shc, and DNA fragmentation was assessed 16 h after suspension. (inset) Immunoblot for Shc with decreased p66^Shc expression. (F) HUVEC were transfected with a vector or CH2, and DNA fragmentation was measured after 16 h of suspension. (G) MDCK cells were plated at different densities and the next day left adherent or forced into suspension. DNA fragmentation at sparse and confluent states was assessed. (inset) Immunoblot for Shc at the indicated levels of confluence. (H) 3T3 cells were plated at different densities and anoikis was assessed as in G. (inset) Immunoblot for Shc at the indicated levels of confluence. (D–H) *, P < 0.001 compared with attached control; †, P < 0.001 compared with floating control. Error bars show mean ± SEM.