Fig 2.

The DD2R transcript is expressed at multiple developmental stages. Reverse transcription (RT) using random hexamers was performed to generate first-strand cDNA (+) from either total RNA (A) or poly(A) mRNA (B–D). In addition, negative controls for each RNA sample were carried out in the absence of reverse transcriptase (−). (A) PCR was performed using the primer pair S4/A4 (as detailed in Supporting Materials and Methods). The products were visualized on an ethidium bromide-stained 1.2% agarose gel. (B–D) The tissue/stage-specific levels of expression of DD2R (B), DmDop1 (C), and actin 5C (D) in adult head (○), adult body (•), pupa (▾), and larva (▴) were quantified by TaqMan analysis as detailed in Materials and Methods. The increase in fluorescence (ΔRn values) was plotted against cycle number (PRISM 3.0, GraphPad). The graphs shown are representative of multiple experiments. No detectable amplification was observed in negative controls (RT−), which were run in parallel for each tissue/transcript examined (data not shown).