Skip to main content
. Author manuscript; available in PMC: 2012 Oct 4.
Published in final edited form as: Cell Metab. 2012 Apr 4;15(4):439–450. doi: 10.1016/j.cmet.2012.02.014

Fig. 6.

Fig. 6

mir-71 facilitates the localization and transcriptional activity of DAF-16 in the intestine of animals lacking germ cells. (A–C) Loss of mir-71 function partially affected the nuclear accumulation of DAF-16::GFP in the intestine of germline-deficient glp-1(e2141) animals. (A) Representative images for each genotype. (B) Loss of mir-71 function does not cause major changes in overall levels of DAF-16::GFP expression in day 2 adults (n = 20). Error bar, standard error of the mean (SEM). (C) Intestinal DAF-16::GFP nuclear accumulation was assessed by measuring the number of nuclei from the images of day 2 adults (low accumulation, <10 nuclei; intermediate accumulation, 10–20 nuclei; high accumulation, >20 nuclei) (n = 78 for each genotype). Error bar, standard error of two biological replicates. All strains contain the daf-16(mu86) allele and the functional muIs109[Pdaf-16::DAF-16::GFP] reporter. (D) Loss of mir-71 function blocked the induction of Psod-3::gfp in day-3 germline-deficient glp-1(e2141) animals (p<0.0001; 40% reduction in mean GFP fluorescence intensity) (wild-type, n=32; mir-71(n4115), n=35; glp-1(e2141), n=40; mir-71(n4115); glp-1(e2141), n=40). (E) Loss of mir-71 function did not affect the expression of DAF-16 targets in intact animals. mRNA levels of DAF-16 target genes sod-3, K04A8.5, gpd-2, dod-8 and nnt-1 were measured by qRT-PCR in wild-type and mir-71(n4115) day-2 germline-intact adults. Loss of mir-71 function did not affect the levels of sod-3, K04A8.5, gpd-2 and dod-8, while it resulted in a 50% increase in the levels of nnt-1 (p<0.05). mRNA levels are relative to wild-type levels. Error bars, standard error of four biological replicates. (F) Loss of mir-71 function reduced the expression of a subset of DAF-16 targets in germline-less animals. mRNA levels of DAF-16 target genes sod-3, K04A8.5, gpd-2, dod-8 and nnt-1 were measured by qRT-PCR in germline-deficient glp-1 and mir-71; glp-1 day-2 adults. Loss of mir-71 function in gelmline-defective glp-1 animals resulted in 50% decrease in the levels of sod-3 (p<0.05) and K04A8.5 (p<0.01) and a small reduction in the levels of gpd-2 (p<0.05). mRNA levels are shown relative to glp-1(e2141) levels. Error bars, standard error of four biological replicates.