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Fig. 6. Mib2 localisation and functional analysis in Drosophila adult
muscle. (A-A''') Mib2 (green) accumulates at the Z band
(Kettin staining, blue) of myofibrils and to lesser an extent in the M band
(absence of actin). (B-D') Electron micrographs of longitudinal
sections of IFM from 3-day-old wild-type (B,B'), 1-day-old (C,C')
and 3-day-old (D,D') 1151-GAL4/UAS-RNAi-mib2 adults. IFMs of
1151-GAL4/UAS-RNAi-mib2 young adults show poorly defined M bands and
irregular Z disks (C'), compared with the wild type (B').
Three-day-old 1151-GAL4/UAS-RNAi-mib2 IFMs lack sarcomeres and
display remains of dense material that probably corresponds to distorted and
split Z bands (D', arrow and arrowhead, respectively). (E-G)
Co-immunoprecipitation experiments showing a physical interaction between Mib2
and nonmuscle myosin. Protein extracts from sqh-GFP thoraces were
immunoprecipitated with anti-Mib2 (E), anti-GFP (F) or anti-nonmuscle Mhc (G)
antibodies, followed by immunoblotting with anti-GFP (E) or anti-Mib2 (F,G).
Note the presence of bands of 47 kDa corresponding to Sqh-GFP (arrowhead in E)
and 115 kDa corresponding to Mib2 (arrowheads in F,G) in the experimental and
their absence in the control P lanes. In E, the weak bands detected in P lanes
correspond to immunoglobulins. Input, immunoprecipitation reaction samples; P,
immunoprecipitated pellets; S, supernatants. Scale bars: 0.5 µm in B; 0.1
µm in B'.
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