12915_2019_646_MOESM3_ESM.pdf (5.12 MB)
Additional file 3: of Enabling cell-type-specific behavioral epigenetics in Drosophila: a modified high-yield INTACT method reveals the impact of social environment on the epigenetic landscape in dopaminergic neurons
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posted on 2019-04-10, 05:00 authored by Pavan Agrawal, Phuong Chung, Ulrike Heberlein, Clement KentComparison of tagged GFP expression in adult Drosophila brain. The INTACT transgene (unc84-2XGFP) was driven in dopaminergic neurons (TH-GAL4) using different copy numbers of the UAS promoter and expression of GFP was compared using the same imaging settings. (A) 3X-UAS- (B) 5X-UAS- and (C) 10X-UAS-unc84-2XGFP. The 3X-UAS-unc84-2XGFP transgene most faithfully reproduced TH-GAL4 expression, while ectopic expression was observed upon further increases of the UAS copy numbers. Dopaminergic neurons were stained with anti-TH antibodies (red), INTACT transgene expression using anti-GFP antibodies (green), and N-cadherin (blue) was used as reference. See Fig. 1b for 3X-UAS-unc84-2XGFP brain imaged at higher intensity. Scale bar is 20âÎźm. (PDF 5243 kb)
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UAS copy numbersGFP expressionadult Drosophila braindopaminergic neuronsUAS promoterEnabling cell-type-specificantibodyscale barDopaminergic neurons10 X-UAS-unc 84-2XGFPINTACT transgene3 X-UAS-unc 84-2XGFP transgene3 X-UAS-unc 84-2XGFP brainanti-GFP antibodiesINTACT method1 bTH-GAL 4 expression3 X-UASectopic expression5 X-UASanti-TH antibodiesimaging settingsPDF 5243 kbdopaminergic neurons Comparisonunc 84-2XGFPcopy numbers
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