Fig. 2

Upregulation of Bmp4 and Shha in the gut of miR-145 morphants. Lateral view of whole-mount in situ expression of bmp4 (A, C) and shha (E, G) shows that both genes have increased expression in miR-145 morphant guts at 96 hpf. Longitudinal sections show that bmp4 (B, D) is expressed in smooth muscle cells (smc; arrowheads) and shha (F, H) is expressed in epithelial cells (ec; arrows). The magnification for B, D, F and H is shown by the scale bar in H. (I) Quantitative PCR shows that knockdown of miR-145 leads to strong up-regulation of bmp4 and shha but not the related gene ihha in miR-145 morphants as compared to control embryos (p<0.01, n=3). (J–K) Increased numbers of pSMAD 1, 5, 8 positive (red; white arrow) smooth muscle cells (green) are found in miR-145 morphants (K) as seen in transverse sections of the gut compared to WT (J), counterstained with DAPI (blue). The scale bar in K represents the magnification for J and K. (L) Graph of the proportion of pSMAD 1, 5, 8 positive SMCs. Significance was determined using the student′s t test (p<0.01)