Figure 2. CMK-1 acts cell-autonomously to regulate daf-7 TGF-β expression in ASI.

(A) Dauers formed by the indicated strains on 80 μg live OP50 and 6 μM ascr#3. Alleles used were: cmk-1(oy21), daf-3(mgDf90), daf-5(e1385), and daf-16(mgDf50). Shown are the averages of ≥3 independent experiments with >65 animals each. Errors are SEM. (B) Representative images of daf-7p::gfp expression in L1 larvae of wild-type or cmk-1(oy21) animals under the indicated conditions. Schematic at top indicates the position of ASI cell body (lateral view); boxed region is shown in panels below. Occasional weak expression is observed in the ADL neurons. Animals were grown with plentiful live OP50 or starved for at least 6 hr in the absence or presence of 1 unit of crude pheromone (see ‘Materials and methods’). White arrowheads indicate cell bodies of ASI. Numbers in bottom left hand corners indicate the percentage of examined larvae that exhibit the shown phenotype; n > 50 each; three independent experiments. Lateral view; scale bar: 10 μm. (C) Scatter plot of fluorescence intensity of daf-7p::gfp expression in ASI in wild-type or cmk-1(oy21) mutants. Median is indicated by a red horizontal line. Animals were grown on ample live OP50 in the absence of exogenous pheromone. Each dot is the fluorescence intensity of a single neuron in a given experiment; n > 60 neurons total each, at least three independent experiments. Promoters driving wild-type cmk-1 cDNA were: ASI—srg-47p; AWC—ceh-36Δp. (D) Dauers formed by shown strains on 80 μg live OP50 and 6 μM ascr#3. The srg-47 promoter was used to drive expression of wild-type daf-7 cDNA in ASI. Shown are the averages of ≥3 independent experiments with >65 animals each. Errors are SEM. Unless indicated otherwise, * and *** indicate different from wild-type at p < 0.05 and p < 0.001, respectively, ^#, ^##, and ^### indicate different from cmk-1 at p < 0.05, 0.01, and 0.001, respectively. (ANOVA and Games-Howell post-hoc test).

DOI: http://dx.doi.org/10.7554/eLife.10110.008