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Fig. 3. Identification of pbl as novel zygotic factor required for mesodermal cell migration. (A-C) Embryos at extended germband stages carrying a synthetic chromosomal deletion produced by C(3)se females crossed to T(3;2)C309/TM3 males (uncovering genomic segment 61 to 68) were stained with anti-EN (brown) and anti-TWI (black) antibodies. The reduction of EN stripes is due to deletion of the hairy locus in this genomic interval and was used as independent marker for the genotype. (A,B) Lateral (A) and ventral (B) views of whole-mount staining. (C) Cross-section demonstrates that the mesoderm cells fail to migrate in these embryos. Wild-type embryos (D) and pbl³ homozygous embryos (E) at stage 11 were stained for EVE (blue) and TWI (brown) protein. In pbl mutants the number of EVE-positive hemisegments is strongly reduced. (F-H) Cell shape changes were analyzed by examining twi::CD2 expression in pbl³ homozygotes; cell shape changes are blocked in pbl mutant embryos. (I,J) F-actin staining (red) of wild-type (I) and pbl³ homozygous embryos. TWI staining is seen in green. F-actin-rich protrusions are indicated with arrowheads in I.

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