Fig. S1

Nup210 role in skeletal muscle growth. Related to Figures 1 and 2.

(A) U2OS cells were transfected with a vector expressing V5-tagged Zebrafish Nup210. Cells were stained using the V5 antibody and the NPC antibody mAb414. Top panel shows a cross-section of the nucleus. Bottom panel shows NPCs on the nuclear surface. Zebrafish Nup210 localizes to NPCs. n ≥ 3 (B) Zebrafish one-cell embryos were injected with control or two different Nup210 morpholinos and slow muscle fibers were stained 48 hours post-fertilization with the F59 antibody. Representative image of a maximum projection of 30-40 sections. (C) Schematic illustration of the myogenic waves in Zebrafish based on Barresi et al (Barresi et al., 2001). During segmentation slow muscle fibers form in the myotomes from adaxial-derived precursors (red, first myogenic wave). When segmentation is complete, at ~24 hpf, new muscle fibers form a different set of muscle precursors and are added to the dorsal and ventral side of the myotome in a process known as stratified hyperplasia (green, second myogenic wave). Sonic Hedgehog (Shh) is required for the first, but not the second, myogenic wave. Thus, the Shh inhibitor cyclopamine inhibits the formation of the early development muscle fibers (red) but not the myofibers that are added post-segmentation (green). (D) Illustration showing the experimental approach to study the role of Nup210 in postsegmentation muscle growth using cyclopamine. (E) Control or Nup210 morphants were stained 48 hpf with the slow muscle F59 antibody and the number of myofibers in myotome 20 were quantified. Bar plots represent mean ± SEM, ∗∗∗∗p ≤ 0.0001, two-tailed Student's t test, n ≥ 3 replicates. Morpholino depletions were performed with n ≥ 50 embryos. n = 10-20 embryos from at least three independent experiments were examined by immufluorescence and quantified.