Skip to main content
. 2022 Apr 29;61:101507. doi: 10.1016/j.molmet.2022.101507

Figure 5.

Figure 5

Nras is a target of miR-29a-3p. (A) Volcano plot demonstrating Nras significantly up-regulated in the arcuate hypothalamic nucleus ARH of DicerCKO-Scrambled mice compared to Control-Scrambled mice (n = 3). (B) Euler diagram identifies Nras as the candidate target gene with the indicated criteria. (C,D) Prediction by Diana-microT-CDS [31] (C) and validation by dual luciferase assay (D) of a highly phylogenetically conserved binding site for miR-29a-3p in the mouse Nras 3′-UTR. (E,F) Functional validation of Nras targeting in vitro assessed by qPCR analyses of expression of miR-29a-3p (E) and Nras (F) upon transfection of HT-22 cells with miR-29a-3p mimic or SpCas9-expressing vector equipped with sgR-1 and -2 targeting miR-29a/b-1 cluster (n = 3). U6 and β-actin were used as the reference genes for normalization. Error bars represent SEM. ∗, p < 0.05; ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001 as assessed by Wald test (A) 1-way ANOVA followed by post-hoc Holm–Sidak pairwise comparison test (D,F) and unpaired two-tailed Student's t-test (E).