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. 2010 Jun 24;6(6):e1001001. doi: 10.1371/journal.pgen.1001001

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Estella, Mann.

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) Time line showing when removing the function of btd and Sp1 affects leg growth (orange shadow). (B) A large Df(btd,Sp1) M+ clone in a T1 leg induced 48–72 hrs AEL and marked by yellow (y) bristles (the clone boundary is indicated by the white dotted line in the left leg). For comparison, the wild type right T1 leg is included in this image. The mutant tissue still maintains leg identity scored by the presence of bracted bristles (arrows, inset). Asterisks mark the segments affected by the clone. The same leg segment nomenclature has been used for all the figures: coxa (cox), trochanter (tro), femur (fem), tibia (tib) and tarsus (tar). (C) Wild type antenna with 1^st antennal segment (a1), 2^nd antennal segment (a2), 3^rd antennal segment (a3) and arista (ar). (D) Df(btd,Sp1) M+ clone induced 48–72 hrs AEL results a strong reduction in size of the a1 and a2 antennal segments, while a3 and the ar are normal. The clone is marked by y. (E) A large btd^XG81 M+ clone in a T2 leg induced 48–72 hrs AEL and marked by y (clone is outlined by white dots) results in a small growth defect in the femur (fe) and tibia (tib), which are also partially fused (arrow). The tarsus (tar), trochanter (tro) and coxa (cox) are unaffected. (F, G, H) The downregulation of Sp1 beginning at the second instar using RNAi affects the growth of the entire leg. Two different Gal4 drivers were used to examine different regions of the leg. (F) The medial part of the leg is strongly reduced in size in dac-Gal4; UAS-Sp1i flies. (G) The distal part of the leg is strongly reduced in size in Dll-Gal4; UAS-Sp1i flies. In this experiment we blocked Gal4 activity prior to the second instar using tub-Gal80^ts. (H) Shows a schematic representation of the expression patterns of the two Gal4 drivers used to downregulate Sp1 function (dac in blue and Dll in red). (I, J) Df(btd,Sp1) M+ clones induced 48–72 hrs AEL and examined in 3^rd instar leg discs. (I) A subset of Df(btd,Sp1) M+ clones (marked by the absence of GFP) show de-repression of Dll in the Dac domain and de-repression of dac in the Dll domain (white arrows). Note that these clones do not affect the expression of Dll and dac in their normal expression domains (green arrows). (J) A subset of Df(btd,Sp1) M+ clones (marked by the absence of GFP) show derepression of hth (arrow). Clones that do not derepress hth are indicated with asterisks.