RNA helicase relative expression during antigenic variation Antigenic variation was induced on a unique VSP-expressing Giardia clone. The primers MK5108 clinical trial used for these determinations were the same as those used for the study of the encystation process. We also designed two additional pairs of primers to determine the relative expression of Giardia Dicer and Argonaute (Ago) transcripts. The relative expression from the thirty one Giardia putative RNA helicases was divided into
earlier (30 min – 1 h) and later (3 – 4 h) up-regulated or down-regulated transcripts. Eight putative RNA helicases were up-regulated after antigenic variation induction, three of them earlier and five later. On the other hand, eight putative RNA helicases were down-regulated, five selleck inhibitor after early induction and three later (Figure 6). Figure 6 Real time quantitative PCR (qPCR) of RNA helicases from G. lamblia during antigenic variation. The relative expressions were calculated after induction of antigenic variation for 30 min – 1 hour
(empty fill pattern) and for 3 to 4 hours (line fill pattern). The relative expression from different helicases was divided into up-regulated (upper panel) and down-regulated (lower panel). Green bars represent significant up-regulation and red bars represent significant down-regulation, gray bars represent no TPCA-1 change in the relative expression. A. Helicases up-regulated during the first 30 min to 1 h. B. Helicases up-regulated at 3 to 4 h. C. Helicases down-regulated during the first 30 min to 1 h. D. Helicases down-regulated at 3 to 4 h. Center inset: relative expression for Giardia Dicer and Argonaute at earlier
or later time points. The ORFs are indicated at the bottom of the graph. The graphs eltoprazine represent the mean of three different measures and the respective standard deviation. A more detailed analysis of the relative expression of the eight putative RNA helicases that were up-regulated after antigenic variation induction showed a slight induction ranging from 1,189 to 1,729 times. In addition, two transcripts from the early up-regulation maintain induction after 3-4 hours. The eight down-regulated putative RNA helicases presented strong down-regulation earlier and significant down-regulation later during antigenic variation. Two of the five early down-regulated RNA helicases maintained low levels of expression after 3-4 h, while one of them was up regulated later. The three transcripts that were down-regulated later presented no significant variations at 30 min-1 h (Figure 6). The relative expression of gDicer presented an early up-regulation that is maintained at later times, while Giardia Ago presented a later up-regulation after 3-4 post induction of antigenic variation (Figure 6, inset).