Transfection and RT-PCR showing reduced ap33 mRNA levels in T. vaginalis trichomonads transfected with the antisense plasmid. Part A shows PCR amplification of the neo coding region in transfected parasites. The ethidium bromide (EtBr)-stained band after electrophoresis in 1% agarose is the PCR product of the neo gene that was amplified using DNA from transfected T. vaginalis parasites. As expected, no PCR product was expected from wt organisms, and a predicted product was obtained from the plasmid used directly during PCR as a control (part A, lane labeled C). Part B gives bands after electrophoresis as in part A showing RT-PCR products for the ap33 S (panel a) and AS transcript (panel b). The RT-PCR product for ap65 and α-tubulin are presented in panels c and d, respectively. Part C illustrates the quantitation of the transcript bands in part B (panel a) for the ap33 transcript in AS-transfected trichomonads compared to S-transfected and wt parasites. The bar graph shows the relative amounts of RT-PCR products for ap33. The amount of wild type ap33 transcript was normalized 100%. Quantification was done following Scion image β program.