Tip60 HAT Activity Mediates APP Induced Lethality and Apoptotic Cell Death in the CNS of a Drosophila Alzheimer's Disease Model
Figure 1
Generation and characterization of dTip60E431Q containing APP or APP-dCT double transgenic flies.
The dominant negative HAT defective lines dTip60E431Q A or dTip60E431Q B (Lorbeck et al., 2011) were introduced into an APP or APP dCT background using standard genetic techniques. (A) Histogram depicting qPCR analysis of exogenous levels of dTip60E431Q in staged F1 second instar larval progeny resulting from a cross between the ubiquitous driver 337 and either dTip60E431Q (lines A and B), APP; dTip60E431Q (lines A and B) or APP dCT; dTip60E431Q (lines A and B). 337-Gal4 crossed to w1118 served as a control. Quantification of the exogenously expressed dTip60E431Q mRNA levels relative to endogenously expressed dTip60 mRNA was done using the comparative CT method with RP49 as internal control as described in (Lorbeck et al, 2011). Asterisks (*) indicate significant fold change between the lines A and B for each genotype with values of p<0.05; n = 3. Error bars represent standard error of the mean. (B) Semiquantitative RT-PCR analysis of APP or APP dCT expression in the different transgenic lines to confirm APP transgene presence. cDNA was prepared as before from staged second instar larvae ubiquitously expressing dTip60E431Q with APP or APP dCT (lines A or B in each case) and PCR amplified using primers that flank a 100 bp region in the N-terminal portion of APP. PCR products were visualized using 2% agarose gel containing ethidium bromide. Staged second instar larvae ubiquitously expressing APP or APP dCT were used as controls.