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. 2012 Aug 9;8(8):e1002878. doi: 10.1371/journal.pgen.1002878

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© 2012 Fasulo et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) Overview of LacI-GFP/LacO tethering assay. The GFP-tagged LacI fusion protein (LacI-GFP) binds 256 tandem LacO sequences inserted at position 60F on the second chromosome allowing the detection of the locus in living cells. The HS83 heat-shock promoter (hs) that drives LacI-GFP is activated at 37°C. (B-E) Live analysis of the LacI-GFP signal at 60F in the polytene chromosomes of third-instar larvae. UAS-dMi-2⁺ 3-3/HS83-LacI-GFP LacO; UAS-dMi-2⁺ 15-1/da-GAL4 GAL80^ts individuals were cultured at 18°C until the middle of the third larval instar, maintained at 18°C (B and D) or shifted to 29°C (C and E) for 27 hours, and heat-shocked at 37°C for 1 hour to induce LacI-GFP expression. Both longitudinal (B and C) and transverse (D and E) views of the lacO array are shown.