Expression of ChAT in Tg(mnx1:GFP) spinal motoneurons. (A) Confocal images of a lateral view of 24 hpf embryos processed with antibody labeling showing ChAT in MN soma and axons (arrows). Arrowheads point to ChAT-mnx1:GFP+ cells. (B–D) Quantification of ChAT and mnx1:GFP labeled cells in 24 hpf Tg(mnx1:GFP) embryos. (B) ChAT+ and (C) mnx1:GFP labeled cell numbers in 100 μm segment in rostral and caudal spinal cord of 24 hpf embryos. (N = 10 embryos; 527 mnx1:GFP+ neurons, 307 ChAT+ neurons). (D) Percentage of ChAT expressing mnx1:GFP neurons in rostral and caudal spinal cord. (N = 7 embryos; 186 mnx1:GFP+ neurons, 172 ChAT+ neurons). (E) Confocal images of a lateral view of a 6-day-old larva processed with antibody labeling showing that ChAT-mnx1:GFP+ cells (arrowheads) are found more in the rostral than caudal spinal cord. (F–H) Quantification of ChAT and mnx1:GFP labeled cells in 6-day-old Tg(mnx1:GFP) larvae. (F) ChAT+ and (G) mnx1:GFP labeled cell numbers in 100 μm segment in the rostral and caudal spinal cord of 6-day-old Tg(mnx1:GFP) larvae. (H) Percentage of ChAT expressing mnx1:GFP neurons in rostral and caudal spinal cord. (N = 6 larvae; 499 mnx1:GFP+ neurons, 381 ChAT+ neurons). All images are oriented with rostral towards the left and dorsal to the top. Scale bar: 20 μm. 2-tailed non-paired Student’s t test; n.s.: not significant; *p < 0.05; **p < 0.005.
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