Figure 2.

Activity-dependent suppression of the proapoptotic gene Puma protects neurons. A, Activity-dependent suppression of the proapoptotic gene Puma. QRT-PCR analysis of Puma expression in cortical neurons treated with the indicated drugs. B, Western blot illustrating regulation of Puma expression at the protein level. C, The Puma promoter is sufficient to confer activity-dependent suppression on a reporter gene. Neurons were transfected with a Puma-luciferase reporter, plus a TK-Renilla control vector. At 24 h after transfection, neurons were stimulated for the indicated times, and firefly luciferase reporter activity was measured, normalized to the Renilla control. *p < 0.05, Student two-tailed t test (n = 4). D, E, Puma expression is sufficient to induce apoptosis in cortical neurons. Neurons were transfected with the indicated vectors, plus a vector encoding eGFP as a transfection marker. Transfected neurons were identified after 10 h, and their fate was monitored at 20 and 30 h. *p < 0.05 (n = 3).