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. Author manuscript; available in PMC: 2017 Aug 14.

Published in final edited form as: Eur J Immunol. 2016 Apr 15;46(6):1490–1503. doi: 10.1002/eji.201545663

(A) The miR-204/211 putative binding sites were predicted on TargetScan using CD5 3′ UTR and the seed region sequences are indicated. Arrows represent the PAS location on CD5 3′ UTR. (B) Conservation analysis of miR-204/211 binding sites performed by alignment of the CD5 3′ UTR sequences of seven mammalian species using the Geneious v4.8 software. Pairwise percentage identities are indicated below. (C and D) qRT-PCR analysis of miR-204 expression in resting and activated human primary T cells and Jurkat cells. Data are shown as mean +SD (n= 3 replicates) and are pooled from 3 independent experiments. *p <0.05; Student’s t-test (E) The two miR-204/211 target sites located in the Luc-pA3 luciferase reporter plasmid were deleted by site-directed mutagenesis originating plasmid Luc-pA3ΔmiR. (F) Luciferase activity was quantified 48 hours after transfection of Jurkat in resting and activated state (using anti-CD3/CD28). Data are shown as mean +SD (n= 3 replicates) and are pooled from 3 independent experiments. ***p < 0.001; unpaired t test with Welch's correction.

(A) The miR-204/211 putative binding sites were predicted on TargetScan using CD5 3′ UTR and the seed region sequences are indicated. Arrows represent the PAS location on CD5 3′ UTR. (B) Conservation analysis of miR-204/211 binding sites performed by alignment of the CD5 3′ UTR sequences of seven mammalian species using the Geneious v4.8 software. Pairwise percentage identities are indicated below. (C and D) qRT-PCR analysis of miR-204 expression in resting and activated human primary T cells and Jurkat cells. Data are shown as mean +SD (n= 3 replicates) and are pooled from 3 independent experiments. *p <0.05; Student’s t-test (E) The two miR-204/211 target sites located in the Luc-pA3 luciferase reporter plasmid were deleted by site-directed mutagenesis originating plasmid Luc-pA3ΔmiR. (F) Luciferase activity was quantified 48 hours after transfection of Jurkat in resting and activated state (using anti-CD3/CD28). Data are shown as mean +SD (n= 3 replicates) and are pooled from 3 independent experiments. ***p < 0.001; unpaired t test with Welch's correction.