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. 2019 Nov 25;9:17449. doi: 10.1038/s41598-019-53868-x

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© The Author(s) 2019

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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miR-29b knockdown decreased mature miR-29a/c expression levels, however did not change miR-29a/c genome sequences. (a) qRT-PCR assay showed that the mature levels of miR-29a and miR-29c were significantly downregulated in clones cas1-1, cas1-2, cas2-1 and cas2-2 compared to px458. The data shown represent the mean change with standard deviation from three times sample collection. Statistical analysis was performed using student t-test. *p < 0.05, **p < 0.01, ****p < 0.0001. (b) Surveyor assay detecting the changes on the nucleotide sequences of mmu-mir-29a and mmu-mir-29c were performed using primers designed spanning the genes locus. mmu-mir-29a displayed no changes among clones; mmu-mir-29c showed an increase in clone cas2-3. (c) qRT-PCR detected the downregulation of mature miR-29a and miR-29c in all four HeLa clones. Data shown represent the mean change with standard deviation from three experiments. Statistical analysis was performed using student t-test. *p < 0.05, ***p < 0.001, ****p < 0.0001. (d) Surveyor assay showed that no significant cleavage occurred on hsa-mir-29a/c sequences. (e) Sanger sequencing displayed no nucleotide changes on the genome locus of hsa-mir-29a and hsa-mir-29c. Surveyor assay products were visualized on Bioanalyser equipment using DNA 1000 chip. Full-length Bioanalyser images for (b,d) are presented in Supplementary Fig. 6.