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. 2022 Apr 29;61:101507. doi: 10.1016/j.molmet.2022.101507

Figure 2.

Figure 2

Loss of miR-29a in the arcuate hypothalamic nucleus during adulthood leads to obesity in mice. (A) Scheme representing a genetic context of the mmu-miR-29a/b-1 cluster and the single guide RNAs (sgR) targeting it. The cutting positions are indicated by red triangles. (B) On-target effectivity of sgR-1 and -2 in HEK293T cell line by split luciferase assay (n = 5). (C) Scheme of the experiment to knock-out the miR-29a/b-1 cluster in the arcuate hypothalamic nucleus (ARH) of adult mice. (D) qPCR analysis of pri-miR-29a/b-1 in the microdissected ARH of miR-29aCKO mice with β-actin used as a reference gene. (E–G) Body weight (E), fat pad weights (F) and microphotographs of hematoxylin and eosin-stained perigonadal adipose tissue (G) in adult miR-29aCKO mice bilaterally injected into 2 coronal planes of ARH by a mixture of adeno-associated viral vectors (rAAVs) equipped with single guide RNAs- and CAG-Cre. n = 7 and 4 (D), 5 and 7 for (E), 5 and 4 for (F) for miR-29aCKO and controls, respectively). Error bars represent SEM. ∗, p < 0.05; ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001 as assessed by unpaired two-tailed Student's t-test (B, D, F) or 2-way ANOVA followed by post-hoc Holm–Sidak pairwise comparison tests (E). Scale bar (μm): 125.