spacer gif spacer gif spacer gif spacer gif ARCHIVE ANNOUNCEMENT! spacer gif
QUICK SEARCH:   [advanced]


spacer gif
     Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents    

Right arrow Help viewing high resolution images
Right arrow Return to article
(Downloading may take up to 30 seconds.
If the slide opens in your browser, select File -> Save As to save it.)
Click on image to view larger version.



Fig. 2. Analysis of rhomboid function during embryogenesis using RNA interference. (A) High concentrations of rhomboid-1 dsRNA were important in maximizing penetrance, with 2 mg/ml dsRNA yielding most penetrant phenocopies. A-C show the percentage of injected embryos that hatched. Essentially all rhomboid-1-injected embryos that did not hatch had missing row 1 denticles, polarity reversal of row 4 denticles, and occasional belt fusions characteristic of the spitz group denticle phenotype. (B) Although dsRNA corresponding to both rhomboid-1 and sprouty caused lethality with strong phenotypic effects, none of the remaining rhomboids yielded any discernible phenotypes when inactivated in wild-type embryos. UN, uninjected embryos; buf, those injected with buffer only. (C) Injection of dsRNAs corresponding to rhomboid-2 and -4 into rhomboid-3 null embryos did not produce lethality, or discernible modification of the denticle phenotype resulting from co-injecting rhomboid-1 dsRNA. Note that co-injecting rhomboid-1 dsRNA with other dsRNAs resulted in decreased lethality because it reduced the concentration of rhomboid-1 dsRNA. Injection of rhomboid-1, or rhomboid-3 rhomboid-1 double mutant embryos resulted in 25% lethality because of the lethal phenotype of mutated rhomboid-1 (present in 25% of embryos derived from a cross between heterozygous parents), but no enhancement of the spitz group denticle phenotype was evident when rhomboid-2 and -4 dsRNA was injected (not shown). (D) Phenotypic analysis of wild-type embryos injected with rhomboid-1 dsRNA (white bars), rhomboid-3 null embryos injected with rhomboid-1 dsRNA (grey bars), and rhomboid-3 null embryos injected with only buffer (black bars). Only embryos injected with buffer displayed wild-type cuticles, while embryos injected with rhomboid-1 dsRNA displayed spitz group cuticle phenotypes. Strikingly, injection of rhomboid-3 null embryos with rhomboid-1 dsRNA produced a marked increase in denticle belt fusions compared to wild-type embryos injected with rhomboid-1 dsRNA.





Right arrow Return to article