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First published online 23 January 2008
doi: 10.1242/dev.016006

Development 135, 813-817 (2008)
Published by The Company of Biologists 2008
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Recruitment of Drosophila Polycomb-group proteins by Polycomblike, a component of a novel protein complex in larvae

Urmi Savla, Judith Benes, Junyu Zhang and Richard S. Jones*

Department of Biological Sciences, Southern Methodist University, Dallas, TX 75275-0376, USA.


Figure 1
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  		Fig. 1. Gel filtration analysis of Pcl and E(z) proteins in Drosophila larval nuclear extracts. Duplicate western blots of larval proteins fractionated on a Superose 6 column were probed with anti-Pcl (top) or anti-E(z) (bottom) antibodies. Elution positions of molecular mass standards are indicated above the appropriate fraction numbers.

 

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  		Fig. 2. Non-reciprocal interdependence of E(z), Pcl and Phol for PRE binding. (A) Schematic of the bxd regulatory region of the Ubx gene. PCR-amplified regions are indicated below. (B-E) Quantitative ChIP results using (B) anti-E(z), (C) anti-Pcl, or (D) anti-Phol antibodies, or (E) no antibodies. Wing imaginal discs were dissected from the following larvae: Oregon R (WT); E(z)61 homozygotes [E(z)61]; PWIZ-Pcl/P{GAL4-da.G32} (Pcl RNAi); phol81A; pho1 homozygotes (phol; pho). (F) Western blot showing Pcl and E(z) levels in Oregon R (1), P{GAL4-da.G32}/+ (2) and PWIZ-Pcl/P{GAL4-da.G32} (3) wing imaginal discs.

 

Figure 3
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  		Fig. 3. Non-reciprocal genome-wide dependence of E(z) on Pcl for chromosome binding. Polytene chromosomes from (A) Oregon R, (B) E(z)61, or (C) pUAST-R57-Pcl/P{GawB}c729 larvae were stained with anti-E(z) or anti-Pcl (red) and anti-IIoSer2 (green). (A) Wild-type distributions of E(z) and Pcl. (B) Inactivation of E(z)61 does not affect chromosome binding by Pcl. (C) Pcl knockdown results in loss of chromosome binding by E(z) (background debris signal is indicated by an arrowhead).

 




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