DEV014027 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. dsRNA-injected embryos were scored into four phenotypic classes. Examples of each class are shown from a representative experiment. (A-C) Embryos scored as wild type. (D-F) Embryos scored as ‘marginally affected’. Effects are equivocal or limited to a single hemisegment. (G-I) ‘Clearly affected’ embryos. Abnormalities consistent with the high-concentration RNAi phenotype are visible in multiple hemisegments. (J-L) ‘Unscorable’ embryos are highly and somewhat variably dysmorphic. This phenotype was seen with all types of injections, including buffer-only controls, and is presumed to reflect damage from the injection process. These embryos were excluded from all subsequent analyses.

• Supplemental Figure S2 -

  Fig. S2. Titration of individual dsRNAs. Each panel shows the results of a single experiment. (A-C) Titration of mib2 dsRNA, in the context of total dsRNA concentrations of 2 mg/ml (A), 100 µg/ml (B) and 500 µg/ml (C). (D-F) Titration of Grip dsRNA at total dsRNA concentrations of 2 mg/ml (D,E) and 500 µg/ml (F). (G,H) Titration of perd dsRNA at total dsRNA concentrations of 2 mg/ml (G) and 20 µg/ml (H). Note the differing effects of 20 µg/ml perd dsRNA in these two experiments. In light of this, subsequent titrations were performed at a constant dsRNA concentration of 500 µg/ml, including mew (I,J) and if (K,L).

• Supplemental Figure S3 -

  Fig. S3. A Perl script to facilitate blind scoring of embryo images. Embryo photographs were computationally scrambled using this simple Perl program, which changes the filenames of all TIFF files in the working directory to code filenames and creates a lookup table (‘key.xls’) to associate these codes with the original, informative filenames. To use with non-TIFF images, simply replace all instances of ‘.tif’ in the code with the appropriate file extension.