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. 2022 Jun 8;13:3180. doi: 10.1038/s41467-022-30778-7

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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. 1 — a The miRNAs miR-1, -133a/b and -206 are encoded in three clusters located on mouse chromosome (Chr.) 1, 2 and 18. The primary sequence of mature miR-1/133a is identical; miR-206 varies from miR-1 only by four bases, miR-133b differs in only one base from miR-133a (red). The seed sequence, essential for miRNA-target interaction, is identical in miR-1/206 and miR-133a/133b, respectively (bold blue). b Relative expression of miR-206 in wild-type (control) and adult miR-1/133a dKO extensor digitorum longus muscles (EDL) and soleus muscles (Sol) analyzed via TaqMan assay (n = 4 ctrl/4 dKO, n = 4 ctrl/4 dKO [males/females, 13–17 weeks], Mann-Whitney-U test, one-tailed, *p = 0.0143, Data are presented as mean values +/− SEM). U6 snRNA served as control. c Percentages of miR-206/133b sKO and miR-1/133a dKO animals at postnatal day 21 (P21) are in accordance to the expected Mendelian distribution (likelihood 25%), while miR-1/206/133 tKO animals were missing at P21. Analysis of E18.5 animals reveals presence of miR-1/206/133 tKO animals at this developmental stage (P21: n = 147 control littermates 48 sKO, n = 100 control littermates/35 dKO, n = 34 control littermates/0 tKO at P21; E18.5: n = 26 control littermates 6 tKO). d Gross morphology of control and miR-1/206/133 tKO pups after caesarean section (control = littermates, scale-bar: 1 cm [E18.5]). e Box-plot of miR-1/206/133 tKO and control body weight (n = 63 control/16 tKO, control = littermates, Mann-Whitney-U test, one-tailed, ns = not significant, Box indicates median and 25th to 75th percentiles, whiskers indicate minimum and maximum [E18.5]). f, g GSEA of miR-1/133/206 tKO compared to miR-1/133a dKO quadriceps muscle transcriptome data reveals enrichment of predicted miR-1/206 and miR-133 targets in miR-1/133/206 tKO (n = 4 dKO/4 tKO, familywise-error rate method (FWER), p = <0.001 (f), p = 0.05 (g) [E18.5]). Source data are provided as a Source Data file.