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. 2020 Oct 2;11:4964. doi: 10.1038/s41467-020-18802-0

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© The Author(s) 2020

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 Differential expression of the potential target genes of PMPs and miR-1915-3p is shown. The selected 9 genes are shown in the bottom panel, which were downregulated consistently and significantly (<0.667-fold changes and p < 0.05) with PMP treatment in gene expression profiling analysis and potentially interacted with miR-1915-3p through bioinformatic analysis (Venn diagram, upper panel). b qPCR analysis of potential target gene expression in the MNC, UT-7, K562 and Meg-01 cells. RHOB was consistently downregulated in all cells detected (normalized to Saline, n = 3 independent experiments per condition). c Quantification of the RHOB protein levels by Western blotting analysis (normalized to Saline, n = 3 independent experiments per condition). Lower panel: Representative Western blotting images of RHOB expression. RHOB was suppressed with PMP treatment. d Upper, putative miR-1915-3p binding sequence in the RHOB 3′-UTR. Middle, the scheme of the luciferase reporter vector with the wild-type miR-1915-3p recognition fragment from the RHOB 3′-UTR or mutant recognition site. miR-1915-3p overexpression decreased the luciferase activity of the reporter containing wild-type binding sequence of RHOB compared with that of the reporter containing the mutant form and the controls (n = 3 independent experiments, one-way ANOVA with Bonferroni’s multiple comparison test, p = 0.0009). e RHOB was downregulated in the K562, Meg-01, UT-7, and MNC cells transiently transfected with miR-1915-3p as evaluated by qPCR (left) and Western blots (right) (normalized to NC, n = 3 independent experiments per condition). f RHOB was upregulated in the K562, UT-7 and Meg-01 cells stably transfected with a miR-1915-3p sponge as evaluated by qPCR (left) and Western blots (right) (normalized to pcDNA3.0, n = 3 independent experiments per condition). g Rhob was downregulated in the ALI mouse BM nucleated cells as evaluated by qPCR (left) and Western blotting (right) (normalized to CON mouse, n = 9 mice). All data are expressed as the mean ± S.D, P-value. Two-tailed, unpaired Student’s t-tests were used unless otherwise specified. Source data are provided as a Source Data file *P < 0.05, **P < 0.01, ***P < 0.001.