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Beatrice Romagnolo, Min Jiang, Moni Kiraly, Carrie Breton, Rebecca Begley, John Wang, Jim Lund, and Stuart K. Kim





fig.1

Figure 1. a. HS::let-60(G12V) hermaphrodite in the L4 stage following heat-shock treatment, showing excess vulval induction. b. Wild-type hermaphrodite in the L4 stage following heat-shock treatment. c. HS::let-60(G12V) hermaphrodite in the L3 stage following heat-shock treatment, showing a Clr phenotype. d. Wild-type hermaphrodite in the L3 stage following heat-shock treatment. Scale bars, 10uM.



fig.2a

Figure 2 a . Hierarchical cluster of predicted heat-shock proteins. Shown are expression profiles of 29 genes encoding Heat-shock proteins in the let-60(G12V) and the wild-type time courses, ordered by hierarchical clustering. Column headings refer to hours after heat-shock. Expression levels at each time point are normalized to expression before heat-shock. Single asterisks indicate genes that show maximum expression changes greater than 2 fold, and double asterisks indicate genes that exhibit more than 4 fold induction. Scale shows level of expression. Identities and expression levels of individual genes are available by clicking the image. You may need to set your web browser at a font of Times New Roman with size 10 to obtain a better image from the GeneExplore.

fig.2b

Figure 2 b. Expression of let-60 mRNA following heat-shock treatment of HS::let-60(G12V) and wild-type strains. Shown are the average expression levels for four to eight repeats at each time point (+/- SD), normalized to expression before heat-shock. The expression of let-60 in the let-60(G12V) time course is significantly greater than in the wild-type time course (Student's t-test, p<0.001).




fig.2c

Figure 2 c. Western blot analysis of MAP kinase activation. Top panel shows staining using monoclonal antibodies specific for the phosphorylated form of mammalian MAP kinase. Bottom panel shows staining using monoclonal antibodies against total mammalian MAP kinase protein. Columns refer to hours following heat-shock treatment in the time courses for HS::let-60(G12V) and wild-type strains. Sizes of molecular weight standards (Kd) are indicated. mpk-1 expresses two forms of MAP kinase protein by alternative splicing [Lackner, 1998], but only one form was stained by the anti-phospho-MAP kinase monoclonal antibody.




fig.3a

Figure 3 a. Pie chart shows the distribution of 297 let-60-regulated genes. Gene annotations were derived from Proteome, and were used to manually place genes into functional classes. Numbers of genes in each class are shown, and do not include 411 genes that encode novel proteins. PC users may click the colored portions of the pie chart to view the gene lists and click the center to view the entire gene list. Mac users can go to page Supplemental Tables to view the gene lists.



fig.3b

Figure 3 b. Hierarchical clustering of 708 let-60(G12V)-regulated genes. Shown are the average expression levels of 708 genes with significant changes in expression levels between the let-60(G12V) and the wild-type time courses. Columns show hours after heat-shock treatment in each of the time courses. Expression levels are normalized to expression before heat-shock treatment. Hierarchical clustering was used to place the genes into groups (A, B and C). Asterisk denotes expression of let-60. Scale shows level of expression. Click the image to explore genes. You may need to set your web browser at a font of Times New Roman with size 10 to obtain a better image from the GeneExplore.



fig.4

Figure 4. Expression profiles of let-60-regulated genes. a. sos-1. b. Transcription factors. c. Ion channels and neurotransmitter receptors. d. Neurotransmitter metabolic proteins. e. G protein-coupled receptors. f. Cell surface glycoproteins. g. Drug resistance proteins. h. Metalloproteases. Genes discussed in the paper are indicated. Scale shows level of expression. Click the image to explore genes. You may need to set your web browser at a font of Times New Roman with size 10 to obtain a better image from the GeneExplore.



fig.5

Figure 5. Shown are the expression of 34 genes in the let-60(G12V) time course, the let-60(n1046) time course compared to the wild-type time course (let-60(n1046)/WT), the let-23 time course compared to the wild-type time course (let-23/WT) and the let-60 time course compared to the let-23(sy1) developmental time courses let-60(n1046)/let-23). These genes showed stronger regulation in the let-60(n1046)/WT time course than in the let-23/WT time course. Scale shows level of expression. Click the image to explore genes. You may need to set your web browser at a font of Times New Roman with size 10 to obtain a better image from the GeneExplore.



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