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Figure 5 | BMC Developmental Biology

Figure 5

From: Genetic and physical interaction of Meis2, Pax3 and Pax7 during dorsal midbrain development

Figure 5

Meis2 containing protein complexes in the mesencephalic vesicle. (A) Co-precipitation of Pax3 and Pax7 with Meis2 in GST-pull down (left panels) and immunoprecipitation experiments using a Meis2 specific antibody (right panels). Upper panel: Western Blot with an antibody directed against Pax7; lower panel: Western Blot against Pax3. (B) Schematic representation of the Meis2 deletions N-terminally fused to GST that were used in (A, C, D, E, and F). (C) Pull down experiments using full length Meis2 (Meis2[1-400]), Meis2 lacking the MEINOX-domain (Meis2[199-400]; ΔMD) or Meis2 lacking the homeodomain (Meis2[1-190]; ΔHD) probed for Pax7. (D) GST-pull down with full length Meis2, ΔMD, ΔHD and GST probed for Pbx1b. (E) Specific DNA complex formation on a 26-bp oligonucleodite and purified GST proteins. Complex formation is competed by specific oligomers at a 10×-molar ratio ('competitor'). The sequence of the 23P-labeled oligomer is indicated below. (F) GST-pull down with ΔN and ΔC probed for Pax3 (upper panel) and Pbx1b (lower panel). (G) Immunoprecipitation using a Pax7-specific antibody probed for Meis2. (H) The same blot as in (G) stripped and re-probed for Otx2. Cyto: cytoplasmic extract; In: input; IP: immunoprecipitate; no Ab: precipitation with protein G sepharose beads omitting the antibody; nucl: nuclear extract; PD: pull down; sup: supernatant; wash: final wash step of the precipitates; WB: Western blot: The asterisks in (A) and (C) mark unspecific bands

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