1. Drosophila tester strains:
a. mwh: The marker multiple wing hairs (mwh, 3-0.3), which is a completely recessive, homozygous viable mutation, is kept in a homozygous mwh strain. The mwh mutation is located near the tip of the left arm of chromosome 3 and in homozygous condition produces multiple trichomes per cell instead of the normally unique trichome.
b. flr3/In(3LR)TM3, ripp sep l(3)89Aa bx34e e BdS: The marker,flare3 (flr3, 3-38.8) is a recessive mutation that affects the shape of the wing hairs, producing a trichome that has the shape of a flare. It is also located on the left arm of the chromosome 3, but in a more proximal position. All three extant mutant alle-les of flr are recessive zygotic lethal. By contrast, homozygous flr cells in the wing imaginal discs are viable and produce mutant trichomes. Because of their zygotic lethality, theses alleles are kept in stocks over balancer chromosomes carrying multiple inversions (TM3).
c. High bioactivation (HB) line: ORR/ORR; flr3/In(3LR)TM3, ripp sep l(3)89Aa bx34e e BdS. The ORR strain has chromosomes 1 and 2 from a DDT-resistant Oregon R(R) line, which are responsible for a high constitutive level of cytochrome P(CYP)6A2 (47,53). This cross improves the performance of the wing SMART in the case of promutagens activated via cytochrome P450-dependent metabolic pathways. More detailed descriptions of the genetic markers and the balancer chromosome can be found in ref. 53.
2. Plastic vials (Carolina Biological Supply; cat. no. CE-17-3120 or similar).
3. Plastic foam vial plugs (Carolina Biological Supply; cat. no. CE-17-3122 or similar).
4. Drosophila Instant Medium (Carolina Biological Supply, Burlington, NC, USA; cat. no. CE-17-3200) or mashed potatoes (Knorr Co. or similar) (see Note 1).
5. Microscope at 400x magnification, bright field.
6. Stereomicroscope at 8x magnification.
7. Water bath.
8. Heating plate (optional).
10. Microscope slides, frosted at one end at 25 x 75 mm).
12. Tweezers (Sigma-Aldrich; cat. no. T5790 or similar).
13. Fine-meshed stainless-steel strainer.
14. Metal cubes (approx 40 mg each) for weights.
16. Faure's solution: 30 mg Gum arabic, 20 mL glycerol, 50 mg chloral hydrate, 50 mL water.
17. Nail polish.
18. Live baker's yeast.
19. Sucrose or sugar (crystalline).
20. Agar-agar (powdered).
21. Powdered cellulose (Merck), used for acute treatment. 3. Methods
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