Fig. 2

MiR-9 suppressed FGF8. a-f View from rhombencephalon onto the MHB (A-F) and lateral brain views (g-p). Left brain halves were electroporated at HH9/10 with miR-9 duplex (a-c), miR-9-pSil (d-f), pCAX-EGFP (g-i), and a single stranded antisense oligonucleotides against the 3’UTR FGF8 sequence (j—l). Right brain halves transfected with miR-9-LNAi (m-p). The untransfected brain half served as control. ISH was employed to view FGF8 mRNA (a,c,d,f,g,i,j,l,m,o,p), EGFP was immunostained with anti GFP (b,c,e,f,h,k,n,o). FGF8 visualized with FAST RED achieved the red fluorescence in (m, n, o, p). a-f Overexpression of miR-9 reduced FGF-8 expression. EGFP overexpression had no influence on FGF8 expression pattern (g-i). Ectopic expression of an antisense oligonucleotide to the 3’UTR binding site for miR-9 of FGF8 enlarged FGF8 expression (j-l), as did the expression of miR-9-LNAi (m-p). Abbreviations: Mes-mesencephalon, MHB-mid-hindbrain boundary, Rh-rhombencephalon