Fig. 2. Orb and Grk are improperly localized in early stage egg chambers in orb³⁰³ and completely absent in orb³⁴³. Ovaries from wild-type and orb mutant females were stained with antibodies to Orb and Grk protein. Genotypes are as indicated. The two panels at the top show the pattern of Orb and Grk protein accumulation in the germarium, and early stages of a wild-type ovariole. Note that Orb protein can be detected in region 2 of the germarium, where it begins concentrating in the presumptive oocyte. In the stage 1 chamber in region 3 and older chambers, much of the Orb protein is in the oocyte at the posterior end. Grk protein can be detected in the stage 1 chamber that has not yet completely budded off from the germarium (left arrow) and in slightly older stages (right arrow) at the posterior of the oocyte. The two sets of panels in the middle show the Orb and Grk protein staining pattern in orb³⁰³ ovarioles. The Orb protein distribution in orb³⁰³ cysts and chambers is abnormal. Unusually high levels of Orb protein are observed in the cysts in region 2 and 3 of the germarium, and this protein appears to be distributed in all germ cells. High levels of uniformly distributed protein can be seen in pseudo-egg chambers just budded off from the germarium, but then the level of Orb protein begins to decline as the aberrant chambers age. Grk also shows an anomalous pattern of accumulation (panels on right). As indicated by the arrows on the left, Grk is expressed prematurely and abnormally high levels of Grk protein are observed in cysts in region 2 of the germarium. High levels of Grk protein are also visible in stage 1 pseudo-egg chambers that have just budded off from the germarium (arrows on right). When the level of Orb protein begins to decline, Grk protein is also reduced. The two panels at the bottom show that there is little or no Orb or Grk protein in orb³⁴³ ovarioles.