Additional file 8: Figure S8..
Attenuation of Snail1 signaling promotes myocyte differentiation and retention of cells in the lateral epithelium. (A-C) Dorsal view of whole-mount segments. (A) A control scrambled siRNA was electroporated into the lateral somites. At 16 hours post-electroporation few labeled cells have differentiated into myofibers. Other cells, while still residing in the lateral DM have lost their epithelial morphology. (B) In the presence of siRNA to Snail1 both maintenance of transfected epithelial cells within the lateral DM as well as premature myogenesis are apparent. (C) Snail1 over-expression rescues the siRNA-Snail1 knock-down phenotype, as fewer fibers and more scattered cells are observed (arrow). (D-F) Transverse sections 40 hours post-electroporation. (D) Control scrambled siRNA. Labeled cells are distributed in myotome (M), sclerotome and approach the cardinal vein (CV). (E) Knocking-down Snail1 maintains cells within the lateral DM as epithelial cells. (F) Over-expression of Snail1 rescues the effect of siRNA-Snail1 knock-down. The lateral DM is depleted of labeled cells and a notable fraction is able to migrate towards target sites (arrows). (F’,F”) High magnification of the inset in F showing the CV exhibiting co-localization of a labeled cell with SM markers. Endothelial cells were visualized with the Qh1 antibody (blue). (G) Quantification (mean ± SEM) of labeled cell distribution (N = 7 for control; N = 7 for Snail1; N = 6 for siRNA Snail1; N = 3 for Snail1 + siRNA Snail1). *P ≤0.05; **P ≤0.01. Bar: (D-F) 50 μm, (F’-F”) 32 μm. es, desmin; DM, dermomyotome; SEM, standard error of the mean; siRNA, small interfering RNA; SMA, smooth muscle actin.
Format: JPEG Size: 2.1MB Download file
Applebaum et al. BMC Biology 2014 12:53 doi:10.1186/s12915-014-0053-9