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Figure 4


Fig. 4. Overexpression of Miple leads to mesoderm spreading defects. Embryos overexpressing Miple-HA under the twist-gal4 driver labeled for Twist (TWI; A), EVE (B), Tinman (TIN; C), or Myosin heavy chain (MHC; D). (E,F) Higher magnification of an embryo overexpressing Miple-HA double-labeled for EVE (E) and MEF2 (F is the merged image of both). Arrowheads indicate uneven mesoderm distribution (A), segments lacking EVE cells (B), heart cells (C), dorsal muscles (D), or a region in which both cardioblast and EVE-positive pericardial cells are missing or mislocalized (E,F). (G) Western analysis of the embryos overexpressing Miple-HA driven by the twist-gal4 driver indicates a specific HA-positive band. Miple-HA from these embryos bound and eluted specifically from a Sepharose-heparin column (third lane from left). (H) Western blot of SR+ cell extract with anti-GFP, -Actin and -HA. The cells were transfected with HOW(L)-HA together with either GFP fused to wild-type miple 3' UTR (gfp-miple 3' UTR) or to miple 3'UTR mutated at the HOW-binding site (gfp-miple 3' UTR*).





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