Fig. 2

Dop1R2 RNAi flies show decreased Dop1R2 transcript levels. a Transcript levels assessed by RT-PCR. RNA from Dop1R2 RNAi and control flies was reverse transcribed, and PCR was performed in triplicate using primer sets corresponding to endogenous Dop1R2 as well as other biogenic amine receptors (such as Dop1R1, shown as a reference). b Dop1R2 transcript levels are significantly decreased in Dop1R2 RNAi flies (genotype: w¹¹¹⁸;UAS-dsDop1R2/+;Act5C-GAL4/+), compared to controls (genotype: w¹¹¹⁸;UAS-dsDop1R2/+;TM6B/+). The average band intensity of the Dop1R2 RNAi PCR product was compared to that of control PCR product for the same gene. Primers corresponding to Dop1R2 as well as other biogenic amine receptors (Oamb, octopamine receptor; Oct-Tyr, tyramine receptor; 5-HT1A, serotonin receptor 1A; Dop1R1, other D1-like Dopamine receptor; Dop2R, D2-like dopamine receptor) and an Actin5C control were used (Additional file 14: Table S1). The difference in PCR band intensity for the assessed genes (in Dop1R2 RNAi vs. control flies) are as follows: Dop1R2 (mRNA, in/out primers): -8.6; Dop1R2 (RNAi construct, in/in primers): 20.9; Oamb: -1.2; Oct-TyrR: 0.5; 5-HT1A: -1.1; Dop1R1: -0.1; Dop2R: 3.0; Act5C: -1.3. Symbols: Dop1R2 (Dopamine 1-like receptor 2, CG18741), Oamb (Octopamine receptor in mushroom bodies, CG3856), Oct-TyrR (Octopamine-Tyramine receptor, CG7485), 5-HT1A (5-hydroxytryptamine (serotonin) receptor 1A, CG16720), Dop1R1 (Dopamine 1-like receptor 1, CG9652), Dop2R (Dopamine 2-like receptor, CG33517), Act5C (Actin 5C, CG4027). Error bars indicated standard variance of the mean for each gene. Significance was determined for the difference in intensity of the RNAi sample PCR band versus the control w¹¹¹⁸ PCR band using a one-sided t-test. * p < 0.05, ** p ≤ 0.01, *** p ≤ 0.001