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Figure 4 | BMC Genomics

Figure 4

From: Computational prediction and experimental validation of evolutionarily conserved microRNA target genes in bilaterian animals

Figure 4

Example of the 3'-UTR reporter plasmid and experimental validation. The 3'-UTR sequences of DLL1, CNN3, LARP4, ELK3, EIF2C4, TAGLN2, and ATP6V1B2 were subcloned into the Xho I/Not I site of the psiCHECKâ„¢-2 vector. CNN3 was chosen as representative of the eight candidates listed above (see Methods section). (B) HeLa cells were cotransfected with each combination of 100 ng of reporter plasmid and the indicated amounts of each miRNA (DLL1, 5 pmol of miR-34 (a); CNN3, 60 pmol of miR-1 (b); LARP4, 20 pmol of miR-1 (c); ELK3, 60 pmol of miR-124 (d); EIF2C4, 60 pmol of let-7 (e), TAGLN2, 5 pmol of miR-1 (f), ATP6V1V2, 60 pmol of miR-1 (g), and LARP4, 60 pmol of let-7 (h)). Colours depict each miRNA: miR-34 (red), miR-1 (blue), miR-124 (orange), let-7 (green), and negative control (black). The relative expression of the luciferase gene was measured 24 h after transfection. The normalized luciferase activity of the control vector was set as 1.0. The data represent the average of three experiments and SDs. * P < 0.01.

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