Fig. 3
Thr33 phosphomimetic mutation blocks Hsp90α1 function in thick filament organization.
DNA construct expressing the myc-tagged T33A/Y35F double, or T33E and T33D single mutant was co-injected with Hsp90α1 ATG-MO into fertilized eggs of zebrafish. The injected embryos were stained with anti-myc (9E10) and anti-MHC (F59) antibodies at 28 hpf. A-C. A mosaic and cell-autonomous pattern of rescue was detected in myofibers expressing the non-phosporylatable T33A/Y35F mutant. D-I. Expression of the phosphomimetic T33D (D-F) or T33E (G-I) mutant failed to rescue the myosin thick filament defect in skeletal muscles. H. Plot showing the number of rescued myofibers in 10–16 individual embryos injected with T33A/Y35F, T33E or T33D mutant. A few fibers showed a partial rescue for T33E. Scale bar = 20 μm.