Fig. 5

WNT asymmetry pathway components WRM-1, LIT-1, and POP-1 are important for PRY-1-mediated expression of miRNAs and seam cell development. (a) Schematic diagram showing the involvement of PRY-1 and other WNT asymmetry pathway components in the specification of seam cell fates. The levels of nuclear POP-1 are high in the anterior cell but low in the posterior cell. (b) RNAi knockdowns of bar-1, pop-1, wrm-1, sys-1 and lit-1 in control N2 and pry-1(mu38) animals. Each data point represents the mean of at least two batches (each batch with at least 30 worms) and error bar represents the STD. Student’s t-test was used to determine the statistical significance: *p < 0.05 (compared to L4440 control). (c) Representative images of control N2 and pry-1(mu38) animals following pop-1 RNAi. The numbers of seam cells are increased in both cases. The phenotype is particularly enhanced in pry-1 mutants (scale bar 0.1 mm). (d) Representative images of two adjacent POP-1::GFP fluorescing cell pairs from V1-V5 seam cell lineages following control and pry-1 RNAi treatments (A-anterior, P- posterior, each dotted line marks a cell pair, scale bar 20 μm). POP-1 asymmetry is disrupted after pry-1 RNAi, resulting in fewer cell pairs having asymmetric localisation of POP-1::GFP in their nuclei. (e, f) qRT-PCR analysis of miRNAs in pop-1(hu9) and pop-1(RNAi) worms at the L1 stage. Similar to pry-1(mu38), all miRNAs, except miR-246, are overexpressed in both pop-1(hu9) (e) and pop-1(RNAi) (f) animals. Each data point represents the mean of two replicates and error bar represents the SEM, *p < 0.05, **p < 0.01