DNA construct expressing the non-phosporylatable T87A mutant or phospho-mimic T87E mutant was co-injected with Hsp90α1 ATG-MO into fertilized eggs of zebrafish. The injected embryos were double 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 all myofibers expressing the non-phosporylatable T87A mutant. D-I. Expression of the phospho-mimic T87E mutant resulted in a varied rescue on myosin thick filament organization. 14% (n = 35) of the T87E expressing myofibers failed to show any sign of rescue in myosin thick filament organization (D-F). In contrast, 39.5% (n = 100) and 46.5% (n = 117) of the T87E expressing slow myofibers showed a partial (not shown) or a full (G-I) rescue, respectively.
J. Plot showing the number of fully rescued myofibers in T87A injected individual embryos. In addition, the numbers of fully, partially or no rescued myofibers were presented in zebrafish embryos co-injected with the T87E mutant. Scale bar = 20 μm.