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First published online November 24, 2004
doi: 10.1242/10.1242/jcs.01549

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Increased expression of Drosophila Su(var)3-7 triggers Su(var)3-9-dependent heterochromatin formation

Department of Zoology and Animal Biology, University of Geneva, 30, quai Ernest-Ansermet, 1211, Geneva 4, Switzerland

View larger version (119K): [in a new window]   Fig. 1. Orcein staining of polytene chromosomes from third instar larvae. (A) Detail of the chromocentre (arrow) from HA:FL4D larvae after daily heat-shocks (hs) or without heat-shocks. (B) Polytene chromosomes from males and females containing a heat-inducible Su(var)3-7 transgene. Homozygous HA:FL4D males and females kept at 18°C (no hs) or submitted to daily heat-shock at 35°C from the second larval instar stage (hs) or three daily heat-shocks from the first instar larval stage (hs+). Arrows indicate the X chromosome.

View larger version (68K): [in a new window]   Fig. 2. Immunodetection of Su(var)3-7 on polytene chromosomes from male and female larvae containing the heat-inducible Su(var)3-7 transgene. (A) Chromosomes after standard fixation. Green, anti-Su(var)3-7; white, DAPI staining. Larvae were kept at 18°C (no hs) or submitted to daily heat-shocks at 35°C (hs). (B) Chromosomes after the same heat-shock conditions, but a milder fixation regime (see Materials and Methods). Arrows indicate the X chromosome.

View larger version (43K): [in a new window]   Fig. 3. Immunodetection of H3-diMeK9 on polytene chromosomes from larvae containing a heat-inducible Su(var)3-7 transgene. Green, anti-dimethyl-H3K9; white, DAPI staining. Homozygous HA:FL4D or HA:FL1A male or female larvae raised at 18°C (no hs) or submitted to daily heat-shocks at 35°C (hs). Arrows indicate the X chromosome.

View larger version (42K): [in a new window]   Fig. 4. Immunodetection of H3-diMeK27 on polytene chromosomes of homozygous HA:FL4D males raised at 18°C (no hs) or submitted to daily heat shocks at 35°C (hs). Arrows indicate the X chromosome.

View larger version (48K): [in a new window]   Fig. 5. Double-immunostaining of homozygous HA:FL4D male chromosomes. Antibodies against HP1 (red) and H3-diMeK9 (green) were used and corresponding DAPI staining is shown. Larvae were raised at 18°C (no hs) or submitted to daily heat-shocks at 35°C (hs). Arrows indicate the X chromosome.

View larger version (49K): [in a new window]   Fig. 6. Immunodetection of H3-diMeK9, HP1 and Su(var)3-7 in HA:FL1A; Su(var)3-906 homozygous null mutant chromosomes submitted to heat-shock. (A) H3-diMeK9 staining (green) and corresponding DAPI staining. (B) Double staining with Su(var)3-7 antibodies (green) and HP1 antibodies (red) and the corresponding DAPI staining.

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