Fig 2. AMX-2 negatively regulates RAS/MAPK signaling. (A) Fine-mapping of QTL1 with ILs. For each IL, the regions containing the Hawaii (black) genome in the Bristol (grey) background are indicated, and the corresponding VIs are plotted below. Black columns indicate the average VI of three independent lines carrying an introgression and gray columns the average VI of three sibling lines without introgression. Dashed boxes indicate the QTL1a and QTL1b sub-regions. (B) Allele-specific effects of
amx-2 RNAi compared to empty vector controls. (C) Two copies of Bristol but not Hawaii
amx-2 rescue the increased VI of
amx-2(
ok1235);
let-60(
n1046gf) double mutants. (D) Epistasis analysis of
amx-2(
ok1235). The dashed line indicates the wild-type VI of 3. (E-H) Expression pattern of a transcriptional Pamx-2::gfp reporter in the pharynx and head neurons (E), the adult vulva (F), the intestine (G) and some rectalcells (H) of L4 larvae. The scale bar is 10um. (I) Tissue-specific
amx-2 RNAi. Knock-down in the intestine but not the vulval cells increases the VI of
let-60(
n1046gf) mutants (J) Quantitative PCR of
amx-2 and
amx-1. Expression levels were normalized to the N2 wild-type Bristol strain. Error bars in (A) to (I) indicate the standard error of the mean and in (J) the standard deviation measured in three independent experiments. The numbers of animals scored are shown inside the columns. *** indicates p<0.001, ** p<0.01, *<0.05 and n.s. p>0.1 in a Student's t-test.