Fig. 2. Expression of PfGRASP in the asexual blood stages. (A) Transcription of Pfgrasp (red) was analysed by real-time RT-PCR using total RNA extracted from tightly synchronised parasites every 8 hours. A stage-specific control was performed using the early transcribed etramp gene (grey). Relative gene expression is shown in bar graphs. This ratio was calculated by comparing the average transcription of Pfgrasp and etramp with transcription of the housekeeping gene actin, which was set to 1. Relative quantification through real-time RT-PCR showed no stage-specific gene regulation for Pfgrasp. (B) Immunoblot analysis of wild-type parasites (3D7). Proteins from synchronised parasite cultures from samples taken every 8 hours, were separated by SDS-PAGE on a 10% gel under reducing conditions. Approximately equal amounts of parasite protein were loaded. Using anti-PfGRASP-specific antibodies, one major 70 kDa band can be detected throughout the asexual life cycle. Positions of molecular size markers (in kDa) are indicated.