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

. 2000 Jul 24;150(2):293–308. doi: 10.1083/jcb.150.2.293

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

© 2000 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

Extra-nuclear Ca²⁺ promotes polyADP-ribosylation of PARP. (a) PolyADP-ribosylation of PARP in crude nuclei isolated from unstimulated neurons, exposed to increasing [Ca²⁺], 1 min before (preincubation) or during [³²P]polyADP-ribosylation (2 min, 25°C). [³²P]polyADP-ribosylated PARP was immunoprecipitated, electroblotted (Western blot), autoradiographed, and immunolabeled by Vic-5 antibody. (b) The curve shows the average increase (±SD) in [³²P]polyADP-ribosylation of PARP (measured by densitometry) versus the increase in extranuclear [Ca²⁺], relative to PARP [³²P]polyADP-ribosylation in extranuclear [Ca²⁺] = 25 nM (see Materials and Methods) (n = 9). (c) Back-[³²P]polyADP-ribosylation of PARP, in nuclei of depolarized or unstimulated neurons treated with Ca²⁺ influx blockers and the permeant chelator BAPTA AM. (Top) Autoradiograms of [³²P]polyADP-ribosylated PARP in crude nuclei of unstimulated neurons (lanes 1, 3, 5, 7, and 9), and neurons depolarized by a 10-min train of repetitive (10 Hz) 30-volt, 0.1 ms pulses (lanes 2, 4, 6, 8, and 10). Cultured neurons were preincubated for 10 min at 25°C with 1 mM CdCl₂ (lanes 3 and 4) or with antagonists of NMDA-glutamate receptors, MK-80l (20 μM; lanes 5 and 6) and APV (500 μM; lanes 7 and 8), or preincubated (30 min, 25°C) with the permeant Ca²⁺ chelator, BAPTA AM (50 μM; lanes 9 and 10). PARP was immunoprecipitated from the nuclear protein extracts of these neurons by N-20 antibody, subjected to SDS-PAGE, electroblotted (Western blot), and autoradiographed. (Bottom) Immunolabeling of [³²P]-polyADP-ribosylated PARP by Vic-5 antibody in the immunoprecipitates (n = 6).