Figure 6

ATF3 is not required for 6-OHDA-induced cell death. Dopaminergic cultures prepared from ATF3 +/+ and -/- mice were treated with 20 μM 6-OHDA. A) 24 hours after treatment, cultures were fixed and stained for TH. B) TH-positive cells were counted and survival expressed as a percent of untreated control. Data were analyzed by Student's t-test (ns). C) Twenty four hours after treatment, cells were fixed and stained for TH and cleaved caspase-3 (a-caspase-3). TH-positive and a-caspase-3 positive cells were counted and the percentage of TH-positive cells that were also a-caspase-3-positive was calculated. Data were analyzed by two-way ANOVA (treatment: ***, p < 0.001; genotype: ns). D) Total RNA was isolated at the indicated times and Puma and 18S rRNA RNA levels were assayed by RT-PCR. Data were analyzed by two-way ANOVA (treatment: ***, p < 0.001, genotype: ns). E) Cells were fixed 24 hours after treatment and stained for TH and CHOP. F) TH-positive and CHOP-positive cells were counted and the percentage of TH-positive cells that were also CHOP-positive was calculated. Data were analyzed by two-way ANOVA (treatment: ***, p < 0.001; genotype: *, p < 0.05).