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. 2005 Aug 10;25(32):7420–7428. doi: 10.1523/JNEUROSCI.0333-05.2005

Figure 3.

Figure 3.

Mediators affecting neurotrophin induced PARP-1 activation. a, Phosphorylation of PLCγ-1 in rat brain cortical neurons in response to stimulation by NGF-related nerve growth factors. Immunolabeling of PLCγ-1 and phosphorylated PLCγ-1 (phos. PLCγ-1; on Tyr783; see Materials and Methods) are presented. b, Changes in polyADP-ribosylation of PARP-1 assayed by the shift in its isoelectric point. PARP-1 was activated in brain cortical slices after a 5 min incubation (at room temperature, 5% O2, 95% CO2) with NGF (100 ng/ml) or NAP (10-15 m), by cAMP (150 μm dbc-AMP), and by PKC activation after a 30 min incubation of cortical slices with PMA (200 nm). NGF-induced PARP-1 activation was suppressed by blockers of PKC [1 μm Bim-I (Bim) or 0.5 μm Gő6976 (Gö)] and by a blocker of CAMK (10 μm KN-93) added 40 min before stimulation with NGF or NAP. cAMP-induced PARP-1 activation was suppressed by the PKA inhibitor H-89 (20 μm; applied for 40 min before stimulation). Nuclei were isolated from cortical slices treated by the indicated compounds. Nuclear proteins were extracted and analyzed by two-dimensional gel electrophoresis. Immunolabeled PARP-1 (MCA1522) in the nuclear protein extracts is displayed for each treatment (n = 3 for each treatment).