A, B65 cells were treated with combinations of 6-OHDA and Bt2cAMP as indicated and subjected to RT-PCR analysis as described under “Experimental Procedures.” Note the decreased levels of BDNF and Bcl-2 mRNA in 6-OHDA-treated cells, which is reversed by the addition of Bt2cAMP. In contrast, there are no changes in the mRNA levels of MKP-3 or β-actin, two genes that lack CRE elements in their promoter regions. Gels are representative of three independent experiments using a Bio-Rad PTC-100 programmable thermal controller. B, for quantitative RT-PCR, cells were treated as above and analyzed by real time RT-PCR using the Roche LightCycler SYBR Green I system. Data were analyzed in relation to β-actin crossing points using the DΔCt method (66). Each graph reflects the average of three independent experiments. *, p < 0.05 versus media-treated controls (UT)/Both (ANOVA and Student’s t test with Bonferroni correction).