DAF-16 and HSP-1 are present in the nuclear fraction after heat shock. A, Western blot of nuclear and cytosolic subcellular fractions of daf-16::gfp adult animals before heat shock, after heat shock, and after 24 h of recovery. Nuclear and cytosolic fractions were solubilized in 200 μl and 600 μl of Laemmli buffer, respectively, and 20 μl of each sample were loaded. Lamin B1 is a nuclear fraction marker, and GAPDH is a cytosolic marker. Shown is a representative assay of four independent experiments. B, densitometric analysis of the amount of DAF-16 present in nuclear (N) or cytosolic (C) fractions. C, densitometric analysis of the amount of HSP-1 present in nuclear (N) or cytosolic (C) fractions. The optical density values for cytosolic fractions were multiplied by 3 to give corrected values. p values correspond to Student's t test from four independent experiments. N, nuclear extract; C, cytosolic extract. D, domain organization of HSP-1/F26D10.3. Predicted NES starting at amino acid position 397 is shown.