FIG 5.

Med1 is differentially required for GATA/Pnr target gene activation. (A to A″) Expression analysis of Pnr target gene ac-sc in control wing discs. The DC-ac-LacZ transgene expression visualized with anti-β-galactosidase (red channel) recapitulates ac-sc expression in the DC cluster. (B to B″) DC-ac-lacZ expression is cell autonomously lost in Med1^O2 mitotic clones (GFP⁻) generated in the wing disc dorsal compartment. (C to C″) Immunofluorescence analysis of the Pnr target gene wg in a control wing disc revealed with anti-Wg antibody (red). (D to D″) Wg protein is still present in Med1^O2 mitotic clones (GFP⁻). Magnifications of the DC and Wg regions are shown in the middle column, with the red channel shown separately in the right column. (E) Functional transcription assay in Drosophila Kc₁₆₇ cells left untransfected or transfected with plasmids expressing Pnr and/or its cofactors Ac and Da and treated with Med1 dsRNA or control GFP dsRNA. The expression of firefly luciferase was standardized to a transfection control, pAc-Rluc. Three independent transfections were performed. Standard deviations are indicated by error bars.