Supplementary Figure 1: MicroRNA represses target proteins without causing mRNA decay. | Nature Structural & Molecular Biology

Supplementary Figure 1: MicroRNA represses target proteins without causing mRNA decay.

From: A novel class of microRNA-recognition elements that function only within open reading frames

Supplementary Figure 1

a,b, Western blotting analysis of miR-20a target genes in response to increasing doses of miR-20a mimic (a) or antagomir (b). c,d, RT-qPCR analysis of miR-20a levels in HeLa cells transfected with miR-20a mimic (c) or antagomir (d). e,f, RT-qPCR analysis of DAPK3 mRNA levels in HeLa cells transfected with miR-20a mimic (e) or antagomir (f). g, Luciferase reporter constructs containing the miR-20a MRE from DAPK3 (native) in Renilla 5’ UTR or mutated to restore base-pairing in the 5’ end (seed) coupled with progressive mismatches (3MM+seed) in the 3’ end. h,i, Results of the luciferase reporter assays (h) and quantification of luciferase mRNA (i). j,k, mRNA levels of luciferase reporters containing individual MREs from 4 indicated genes that function in both CDS and 3’UTR (j) or those that function only in CDS (k), as shown in main Fig. 2e,f. l,m, Luciferase activities from the reporters containing individual MREs from 4 indicated genes that function only in CDS in response to co-transfected Let-7b mimic (l) or antagomir (m). NC: Non-specific oligo control; SC: Scrambled RNA oligo control. n, Western blotting analysis of three additional Let-7b target genes. The three gene products were analyzed in HeLa cells transfected with non-specific RNA oligonucleotide control (NC), Let-7b mimic, or antagomir, as indicated. Actin served as loading control. The data in c,d,e,f,h,i,j,k,l,m were based on 3 independent experiments. *p<0.05; **p<0.01; ***p<0.001 (Student’s t-test).

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