Fig. 2. Deletion of individual neuronal regulatory elements eliminates expression in the corresponding neurons, without eliminating expression of a reporter driven by the same element. All embryos are in a Df(2R)eve mutant background, and carry the rescue transgene indicated on the left of each row (see Fig. 1 and text for details). All are oriented with anterior towards the left. Two different focal planes of the same embryo, stained with anti-Eve, are shown in the first two columns: the focal plane of the RP2 and a/pCC neurons in A,C,F,I; and that of the EL and U/CQ neurons in B,D,G,J. In all panels, black arrows indicate the positions of RP2 (left arrow) and pCC (right arrow) neurons, arrowheads indicate the positions of U/CQ neurons, and open arrows indicate the positions of EL neurons. (A,B) Eve expression from the complete (`wild type') rescue construct. (C,D) Eve expression from RP2A. Note that there is no detectable Eve expression in RP2 and a/pCC neurons. The positions of RP2 and pCC, which do not overlap with those of U/CQ neurons, are indicated by arrows; aCC is also negative for Eve staining, but its position, immediately anterior to pCC, overlaps with that of a U/CQ, which is just out of focus in C. Eve expression in U/CQ (arrowhead) and EL neurons (open arrow) is normal. (E) ß-Gal expression (brown) driven by the RP2+a/pCC regulatory element in the same neurons where Eve is missing. Note that the element is still active in the absence of Eve (there is no black anti-Eve staining visible in this focal plane). (F,G) Eve expression from U/CQ. (H) ß-gal expression (brown) driven by the U/CQ element in the eve^– neurons, and Eve expression (black) from U/CQ. (I,J) Eve expression from EL. (K) ß-gal expression (brown) driven by the EL element in the eve^– neurons, and Eve expression from EL (black). Scale bar: 50 µm.

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