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. 2011 Oct 18;5(10):e1338. doi: 10.1371/journal.pntd.0001338

Figure 8. Experimental verification of potential target(s) for miR5.

Figure 8

(A) The four genes containing potential miR5 target sites chosen for further test. (B) Two copies of each potential miR5 target site separated by a 10 nt spacer were cloned to the 3′-UTR of Rluc gene to generate four constructs: RL-2TS1, 2, 3 and 4. A Luciferase transcript carrying at the 3′-UTR a double miR4 target sites from another VSP (GL50803_36493) [21], unrelated to that of miR5, was included as a control (RL-2TS-miR4). The in vitro transcripts (4 µg) with or without 1 µg of synthetic miR5 were each introduced into Giardia via transfection and the subsequent Luciferase expression was monitored. The results and standard deviations were from three independent transfection experiments. The p-values indicated were calculated by two-tailed Student's t-test.