Fgf signal suppresses gsx2 expression and IO neuronal development. (A-D) 3 dpf control (fgf8a+/+) and fgf8ati282a/ti282a mafbaGFF;UAS:GFP;gsx2:RFP larvae that received an injection of control (Ctrl) MO and fgf3 MO, respectively, were stained using anti-RFP (magenta) and anti-GFP (green) antibodies. (E-H) mafbaGFF;UAS:GFP;gsx2:RFP were treated with 1% DMSO (E,F, n=4) or 200 µM SU5402 (G,H, n=4) from 6 to 22 hpf, fixed at 3 dpf, and stained using anti-RFP (magenta) and anti-GFP (green) antibodies. (I,J) Length of the gsx2:RFP+ hindbrain region (gsx2 expression) in fgf8a+/+; Ctrl MO (n=9) and fgf8ati282a/ti282a; fgf3MO (n=6) larvae (I), and in DMSO- (n=4) or 200 μM SU5402-treated (n=4) larvae (J). (K-P) Expression of GFP (green) and Pvalb7 (magenta) in control (fgf8a+/+) and fgf8ati282a/ti282a 28C;UAS:GFP larvae that received an injection of control MO and fgf3 MO (fgf3MO), respectively. (Q-V) 28C;UAS:GFP were treated with 1% DMSO (Q-S, n=6) or 5.0 µM SU5402 (T-V, n=3) from 6 to 22 hpf, fixed at 3 dpf, and stained using anti-Pvalb7 (magenta) and anti-GFP (green) antibodies. (L,O,R,U) Higher magnification of the boxes in K,N,Q,T. (W) Number of 28C;UAS:GFP+ IO neurons in fgf8a+/+;Ctrl MO (n=6) and fgf8ati282a/ti282a; fgf3 MO (n=5) larvae. (X) Number of 28C;UAS:GFP+ IO neurons in DMSO- (n=6) or 5.0 μM SU5402-treated (n=3) larvae. r5-6, rhombomeres 5 and 6. *P<0.05; **P<0.01; ***P<0.001 (Student's t-test for J and X, Welch's t-test for I and W). Data are mean±s.e.m. with individual values indicated. Scale bars: 100 µm in A (applies to A-D) and E (applies to E-H); 50 µm in K (applies to K,N,Q,T) and M (applies to M,P,S,V); 20 µm in L (applies to L,O,R,U).
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