DEV009373 Supplementary Material

Files in this Data Supplement:

• Supplemental Table S1 -

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• Supplemental Table S2 -

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• Supplemental Table S3 -

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• Supplemental Table S4 -

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• Supplemental Table S5 -

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• Supplemental Figure S1 -

  Fig. S1. met-1 and met-2 mutants appear to have approximately normal levels of histone H3. (A) The levels of histone H3 as assayed using quantitative western blots are shown. Levels of histone H3 were normalized to levels of histone H2A. Relative levels of histone H3 in met-1(n4337) (white bar) and met-2(n4256) (gray bar) mutants were normalized to relative levels of histone H3 of the wild type (black bar). Normalized units of fluorescence and standard deviations are shown. (B) The levels of histone H3 as assayed using quantitative western blots are shown. Levels of histone H3 were normalized to levels of tubulin. Relative levels of histone H3 in met-1(n4337) (white bar) and met-2(n4256) (gray bar) mutants were normalized to relative levels of histone H3 of the wild type (black bar). Normalized units of fluorescence and standard deviations are shown.

• Supplemental Figure S2 -

  Fig. S2. The histone H3K9 and H3K36 trimethylation antisera are specific. Peptides from histone H3 trimethylated at lysine 9 or lysine 36 (Upstate) were probed with the antisera we used to recognize levels of histone trimethylation in vivo. 0-25 μg of each peptide was blotted. The top row of each blot is the peptide with lysine 9 trimethylated; the bottom row is the peptide with lysine 36 trimethylated. The top blot was probed with antisera to histone H3K9 trimethylation (Upstate), the bottom blot with antisera to histone H3K36 trimethylation (Abcam).

• Supplemental Figure S3 -

  Fig. S3. met-1; met-2 double mutants have a mortal germline defect. Seven lines of met-1(n433); met-2(n4256) animals were scored for sterility. The generation at which each line became 100% sterile is shown.

• Supplemental Figure S4 -

  Fig. S4. The met genes and the hpl genes act redundantly to prevent sensitization to RNAi by feeding. After exposure of animals to cel-1 RNAi, the number of arrested L2 larvae was scored in at least three independent experiments. The average percent of L2 arrested larvae is shown. Error bars indicate standard deviations.

• Supplemental Figure S5 -

  Fig. S5. Quantitative western blots. Western blots of protein extracts from wild-type, met-1 and met-2 embryos. Each sample was run in four adjacent lanes. The Cy5-labeled secondary antibodies (green) recognized trimethylated H3K4 (top) or trimethylated H3K9 (bottom). The Cy3-labeled secondary antibodies (red) recognized histone H3 in both blots. A yellow signal was detected when levels of histone H3 trimethylation were roughly equal to levels of histone H3.