Fig. 1. Anti-Medea antiserum is specific. (A) Anti-Medea detected two classes of embryos from a mating of females bearing germline clones of the null allele Med¹³ with Med¹³–/+ males. One class, zygotic heterozygotes (M^–Z⁺), showed variable subcellular localization of the antigen, cytoplasmic in ventral and lateral regions, and nuclear in many dorsal cells. The other class lacked detectable staining, the zygotic null homozygotes (M^–Z^–). (B) Western analysis of larval extracts with affinity-purified anti-Medea antiserum, and actin as a control. Wild-type larvae (WT) and Mad mutant larvae (Mad¹⁰/Mad¹²) give a similar pattern of strongly staining bands of approximately 97 kDa (arrowhead in B-D), 55 kDa and 47 kDa. These bands were missing in protein extracts from Medea mutant larvae (Med¹/Df Med). (C,D) Comparison of steady state Medea levels in embryo extracts using Western blot analysis, with tubulin as a control. (C) WT versus embryos from cactus^PD74 mothers (cact), to assess the contribution of dorsal tissues to total Medea levels. Total Medea is not decreased when dorsal tissues are absent. (D) WT versus globally increased BMP signaling through either constitutively active (CA) receptor, Sax (nos>Gal4;UAS>saxA) or Tkv (nos>Gal4; UAS>tkvA). Steady state Medea levels are unperturbed by hyperactivation of either BMP receptor.

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