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. 2017 Oct;58(12):5368–5377. doi: 10.1167/iovs.17-22410

Figure 2.

Figure 2

The impact of SNP rs2273626 in the seed sequence of miR-4707 on miRNA production and targeting. (A) The figure shows the predicted hairpin structure of miR-4707 containing rs2273626, which was associated with VCDR and cup area. The mature miRNA sequences (3p and 5p) are shown in red and the position of variants is depicted by an arrow. To examine the effect of rs2273626 on the miR-4707 expression level, HEK293 cells were transfected with GFP-miRNA transcripts containing either the minor allele T or the major allele G. The levels of mature miRNA relative to GFP transcript levels were calculated. (B) Luciferase reporter assays indicating miR-4707-3p–mediated repression of CARD10. HEK293 cells were cotransfected with CARD10 3′UTR luciferase reporter vector and GFP-miRNA transcripts containing either the minor allele T or the major allele G. This experiment indicates a significant difference (P = 0.04) between the relative luciferase activity of the CARD10 3′UTR construct in the presence of miR-4707-3p containing the major allele and the minor allele. Our results suggest that rs2273626 diminishes the regulatory interaction between miR-4707-3p and CARD10, resulting in increased CARD10 levels. All experiments were performed in triplicates and repeated at least three times. Error bars represent standard deviation (SD). NS, nonsignificant.