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. 2017 Feb 16;8:14395. doi: 10.1038/ncomms14395

Figure 1. miR-17 is upregulated in kidney cysts of mouse and human ADPKD.

Figure 1

Nanostring nCounter microarray analysis was performed to compare miRNA expression patterns between 10-day-old wild type (n=3) and Pkd1-KO kidneys (n=3), and 21-day-old wild type (n=3) and Pkd2-KO kidneys (n=3). (a,b) The differentially expressed miRNA families in Pkd1-KO and Pkd2-KO are shown. The red circles mark Log2 fold change when only differentially expressed family members are included. The stacked bars mark Log2 fold change when all expressed family members irrespective of statistical significance are included. The values in parenthesis indicate the percentage contribution of each miRNA family to the total miRNA pool in cystic kidneys. The miR-17 family is highlighted in red. (c) Dysregulated miRNAs exhibited correlated expression pattern (R2=0.87) between Pkd1-KO and Pkd2-KO kidneys suggesting that a common set of aberrantly expressed miRNAs may underlie ADPKD pathogenesis. (d,e) Q-PCR analysis demonstrated that miR-17 is upregulated at cystic stages (P7 & P10 for Pkd1-KO and P21 & P28 for Pkd2-KO) but not at pre-cystic stages (P1 for Pkd1-KO and P14 for Pkd2-KO). In situ hybridization (ISH) was performed using an LNA-modified anti-miR-17 probe. The kidney sections were counterstained with nuclear fast red to mark nuclei. (f) Representative ISH images of kidney sections from wild type, Pkd1F/RCSKO, Pkd2-KO, Pkd2-miR-17∼92KO mice (negative control) and miR-17∼92Tg (positive control) are shown. Expression of miR-17 (blue) was increased in cysts (cy) of Pkd1F/RCSKO and Pkd2-KO compared with renal tubules of wild-type mice. miR-17 expression was abolished in cysts of Pkd2-miR-17∼92KO mice, whereas its expression was induced in renal tubules of miR-17∼92Tg mice, indicating that the in situ probe specifically detects miR-17. (g) ISH was performed on kidney sections from four NHK and ADPKD samples. Representative ISH images from one NHK and ADPKD sample are shown. miR-17 expression was not detected by ISH in NHK kidney sections. The percentage of miR-17-positive cysts per kidney section from four human ADPKD patients (#6–9) is shown in the graph. The total number of cysts per section is shown above each bar. P indicates postnatal day. Error bars indicate s.e.m. * indicates P<0.05, ns indicates P>0.05. Student's unpaired t-test (d,e). Scale bars, 50 μm (f) and 20 μm (g).