Figure 3.
daf-16 Does Not Function Downstream to
daf-2/Nutritional Cues to Regulate Zone 1 MPK-1 Activation and Oocyte Development.Dissected adult hermaphrodite germlines stained for DNA (DAPI, white), dpMPK-1 (red), or REC-8 (green).(A and B)
daf-16(
mu86) germlines on either fed (A) or starved (B) conditions.
daf-16 germlines from fed animals reveal normal dpMPK-1 in zone 1 and oocyte development (A) but reduced dpMPK-1 in zone 1 and decreased oocyte production in the starved condition (B). RNAi analysis was performed in triplicate, and 50 germlines were analyzed for each genotype. The starvation experiment was performed five times, and 20-25 germlines were analyzed each time.(C) Whole-mount DIC and GFP analysis of muIs61, DAF-16::GFP animals. Arrows indicate nuclear staining of DAF-16::GFP in somatic gonadal sheath cells.(D and E) Germlines from
rrf-1;
daf-2 animals at the restrictive temperature with gfp (D) or
daf-16 (E) RNAi treatment. In (E), reduction of
daf-16 in
daf-2 mutant animals results in restoration of the mitotic proliferative germ cells (green arrow) but does not rescue PP defects, stalled oocyte development, or MPK-1 activation in zone 1.(F) Germlines from
daf-16;
daf-2 double mutant animals have normal mitotic zone development and dpMPK-1. ++. nonspecific signal from the intestine. Scale bar, 20 um.