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. Author manuscript; available in PMC: 2019 Sep 3.
Published in final edited form as: Dev Cell. 2016 Dec 29;40(1):95–103. doi: 10.1016/j.devcel.2016.12.001

Figure 3. miR-133b-3p is enriched in exosomes but its primary transcript is only expressed in the mesenchyme while the mature miRNA is detected in both epithelium and mesenchyme.

Figure 3.

(a) Exosomal miRNAs are a subset of SMG miRNAs. N = 3. (b) miR-133a-3p, miR-133b-3p, and miR-409–3p were relatively more abundant in the RNA isolated from the exosome fraction than SMGs. miR-200c-3p is an epithelial-related miRNA. Student’s t-test, * P< 0.05, ** P< 0.01, *** P< 0.001. N = 3. (c) TaqMan qPCR of the primary and mature forms of exosomal miRNAs (miR-133a-3p, miR-409–3p) and epithelial-expressed miR-200c-3p shows that primary and mature transcripts were detected in both epithelium and mesenchyme. N = 3. (d) Expression analysis by TaqMan qPCR of primary and mature forms of miR-133b-3p in isolated epithelium and mesenchyme. Images are epithelium and mesenchyme at embryo day 12, 13, and 13.5. The primary transcript of miR-133b-3p was not detected (ND) in epithelium, although the mature form was found in both epithelium and mesenchyme. (e) Primary miR-133–3p is not detected in cultured epithelium but its mesenchymal expression increases. Analysis of primary and mature miRNA expression, normalized to time 0 (dotted line), in isolated epithelium and mesenchyme cultured separately for 24 h. (f) Brefeldin-A (1ng/ml) treatment of SMGs for 6 h, which reduces exosome secretion, decreases mature miR-133b-3p expression in the epithelium by ~80 %. N = 3. All graphs are means ± s.d.