Fig. 1. P1D9 monoclonal antibody is specific for Rho1 protein in vitro and in vivo. (A) Western analysis of Drosophila Rho family GST fusion proteins hybridized with -GST (top) and P1D9 monoclonal (bottom) antiserum. (B) Western analysis of whole cell (wc) and nuclear (n) lysates prepared from 0- to 2-hour and 0- to 12-hour embryos, respectively. P1D9 recognizes a single major band in whole cell but not nuclear lysates. This band is not detected when primary antibody is omitted. (C,D) P1D9 staining of a stage 14 embryo homozygous for a deficiency that includes the Rho1 locus (C) relative to a sibling control (D). The relative intensity of the staining in these embryos can be directly compared, as both embryos were photographed in the same visual field. (E-J) P1D9 recognizes ectopically expressed Rho1 (E), but not Rac1 (G) or Cdc42 (I). Each protein was overexpressed in the Engrailed domain, as highlighted by green fluorescent protein expression (F,H,J), and embryos were examined at stage 14. For all embryos, anterior is left and dorsal is up. Scale bars: 50 µm.

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