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. 2022 Sep 29;13:5733. doi: 10.1038/s41467-022-33367-w

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 5 — a–b Cumulative frequency distribution of mRNA changes for predicted targets of miR-7 (a) or miR-16 (b) in Sim1-cre;LSL-tdTomato;mir-7^fl/fl versus Sim1-cre;LSL-tdTomato RNA sequencing data. Targets are binned by cumulative weighted context ++ score (cs + +) and compared to all expressed genes. Number of genes per bin for miR-7: black, 16830; blue, 225; orange, 149; red, 112. Number of genes per bin for miR-16: black, 16830; blue, 332; orange, 408; red, 425. c log2(fold change) and mean FPKM (fragments per kilobase of transcript per million fragments mapped) of all predicted conserved miR-7 target genes in Sim1-cre;LSL-tdTomato;mir-7^fl/fl versus Sim1-cre;LSL-tdTomato RNA sequencing data. Targets with >1.3-fold upregulation are shown in red. d Expression (relative to Sim1-cre;LSL-tdTomato) of upregulated targets in Sim1-cre;LSL-tdTomato;mir-7^fl/fl versus Sim1-cre;LSL-tdTomato RNA sequencing data (n = 4 samples, with 4 mice pooled per sample). e Strategy of Cre-dependent target overexpression using a pAAV-FLEx vector. f V5 immunofluorescence in the PVN of WT (representative of 3 mice) and Sim1-cre mice (representative of 3 mice), 4 months after injection with AAV-Snca (scale bar: 250 μm). Green, V5; Blue, Hoechst. g–h Body weight of HFD-fed female WT and Sim1-cre mice injected with AAV-Snca (n = 7 and 9 animals) (g) or AAV-Igsf8 (n = 7 and 9 animals) (h). i Luciferase assay of cells expressing the WT or mutated 3’UTR of Igsf8, with or without miR-7 overexpression (n = 4; representative of two independent experiments). j–m Plasma insulin (j–k) and IGF-1 (l–m) in WT and Sim1-cre mice injected with AAV-Snca (n = 7 and 9 animals) (j, l) or AAV-Igsf8 (n = 7 and 9 animals) (k, m). Data are presented as mean ± SD, where error bars are present. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; no asterisk or n.s. indicates P > 0.05; Kruskal-Wallis test with Dunn’s multiple comparisons test (a, b), 2-way repeated measures ANOVA (g, h; genotype-by-time interaction), 2-way ANOVA with Sidak’s multiple comparisons test (i), or two-tailed t test (j–m). **q < 0.01; ****q < 0.0001; quasi-likelihood (QL) differential expression test (c, d). Source data are provided as a Source Data file.