miR-10a-5p and miR-10b-5p restore colonic motility in aged mice

Figure 2 Expression of miR-10a-5p and miR-10b-5p decreases with age. A: RNA sequence and structure of mouse miR-10a and 10b precursors encoding homologous miR-10a-5p and miR-10b-5p. One nucleotide, U or A differing between miR-10a-5p and miR-10b-5p, is circled; B and C: Expression of miR-10a-5p and miR-10b-5p in the whole gastrointestinal tissue at embryonic stage 18 (E18) and the whole blood from 7 days (7D) to 20 months across different age groups of C57BL/6J mice, measured by qPCR. Data were presented as mean ± SD. n ≥ 3 per group. ^aP < 0.05; ^bP < 0.01; D: QPCR amplicons of miR-10a-5p and miR-10b-5p electrophoresed on a 2% agarose gel. NTC stands for a non-template control. A DNA ladder was loaded on each side the gel; E: Construction of a TOPO^®TA based expression vector cloned with miR-10a-5p or miR-10b-5p PCR products. A primer set, M13 forward and reverse primers, used to amplify PCR products (289 bp) is shown. Restriction enzyme sites for SfcI and Sau3AI are indicated; F: Colony PCR products electrophoresed on 2% agarose gels; G: PCR-amplified products digested with Sfcl and Sau3AI electrophoresed on a 1% agarose gel; H: MiR-10a-5p and miR-10b-5p sequencing chromatograms. One nucleotide, T or A differing between miR-10a-5p and miR-10b-5p, is circled.