Caenorhabditis elegans HCF-1 Functions in Longevity Maintenance as a DAF-16 Regulator
Figure 5
Loss of hcf-1 Promotes the DAF-16 Transcriptional Regulation of Several Target Genes
(A) The expression of sod-3, mtl-1, and F21F3.3 was elevated and that of C32H11.4 was repressed in the hcf-1 mutants. The elevated expression of sod-3 and mtl-1 in the hcf-1 mutants was completely dependent on daf-16; that of F21F3.3 and C32H11.4 was partially dependent on daf-16. *, p < 0.05 when compared to wild-type (wt). **, p < 0.05 when compared to hcf-1(ok559).
(B) The expression of sod-3 and mtl-1 in the daf-2(e1370);hcf-1(pk924) double mutant showed synergistic up-regulation when compared to the expression in either hcf-1(pk924) or daf-2(e1370) single mutant. *, p < 0.05 when compared to wild-type (wt). **, p < 0.05 when compared to daf-2(e1370). The quantitative data are summarized in Tables 3–5.
The RNA levels of sod-3, mtl-1, F21F3.3, and C32H11.4 were quantified using qRT-PCR and normalized to the internal control act-1. The data for at least three independent experiments were pooled, and the mean normalized RNA level and SEM for each gene in the indicated strains are shown. The mean normalized RNA level for each gene in wt worms was set as 1. p-Value was calculated using Student's t-test. For sod-3 or mtl-1 expression, we analyzed for a synergistic effect in daf-2(e1370);hcf-1(pk924) compared to daf-2(e1370) or hcf-1(pk924) using two-way ANOVA analysis.