Figure 7.

EcR signaling is required for miR-318 expression. (A–B′′) miR-318 expression is absent in the follicle cells overexpressing EcR^DN. Shown are confocal images of stage-10B egg chambers from mir-318^Δ1/+ heterozygous female flies with control or EcR^DN overexpression clones in the follicle cells. GFP expression reflects miR-318 expression in the mir-318^Δ1 allele as described previously. Clones were marked by the presence of RFP. Samples were stained with anti-GFP (green), anti-RFP (red) and DAPI (blue). Bar, 50 um. (C) Predicted EcRE sequence in the mir-318 regulatory region is shown relative to the EcRE consensus sequence. Mismatches are underlined. The position is shown relative to the mir-318 stem loop region. (D) Luciferase activity assays of wild-type and mutated mir-318 regulatory region luciferase reporters upon 20E treatment in S2 cells. Fold changes were measured as luciferase activity of 20E-treated vs. untreated samples. Control samples were cotransfected with the empty vector. Error bars represent standard deviation from four independent experiments. P < 0.05.