Fig. 4
Expression profiles of the C. sinensis GSTo1 and CsGSTo2 according to developmental stages. a Total RNAs (1 μg) extracted from each of the developmental stages were reverse transcribed as indicated on top. The mRNA transcripts of CsGSTo1 and 2 amplified by a semiquantitative RT-PCR were analyzed by 2 % agarose gels with ethidium bromide staining. C. sinensis tubulin gene (CsTub), which was shown to be constitutively expressed throughout developmental stages, was used as a control. The reaction mixture that did not contain reverse transcriptase during synthesis of the first single strand cDNA was used as a negative control. Abbreviations: Egg, C. sinensis egg; MC, metacercaria; 1 wk, 1-week-old juvenile; 2 wk, 2-week-old juvenile; 3 wk, 3-week-old immature; 4 wk, 4-week-old-mature C. sinensis. b Alteration of CsGSTo transcripts by qRT-PCR. The mRNA transcripts in each of the RNA samples (200 ng) were reverse-transcribed and the resulting cDNAs were employed in qRT-PCR as templates. The fold increase was calculated by differences in threshold cycles (ΔΔCT) of the CsGSTo1 and 2 among different developmental stages. CsTub gene was used as normalization control. c Expressional changes of CsGSTo1 and 2 proteins determined by immunoblotting probed with anti-rCsGSTo1 and 2. C. sinensis tubulin (CsTub) was employed as a control. Each lane contained 100 ng protein