Fig. 1. GLH-1 levels are increased in C. elegans kgb-1 mutants. (A) N2 (wild-type) and kgb-1 worms grown at 20°C for 5 days beyond (d>) the L4 stage were analyzed by western blot (1000 worms in the wild-type lane and 800 worms in the kgb-1 lane), examining GLH-1 and GLH-4; ß-tubulin was a loading control. (B) Relative levels of GLH-1 in kgb-1 and N2 worms were compared for 1, 3 and 5 d>L4 at 20°C, with GLH-1 levels for N2 worms at 1d>L4 set to 1 and other levels determined relative to this. Values are the average of three experiments, with the standard error shown. ^*, differences in GLH-1 between kgb-1 and N2 are statistically significant (P<0.05). Beneath, a representative blot with a ß-tubulin loading control is shown. For B-D, 600 worms were loaded per lane. (C) N2 and kgb-1 worms grown at 26°C for 3.5d>L4 were analyzed by western blot to examine GLH-1 and GLH-4 levels. (D) GLH-1 levels were quantified for N2 worms and kgb-1 mutants grown at 26°C for 1, 2 and 3d>L4. ^*, GLH-1 differs significantly between N2 and kgb-1 animals (P<0.05). ß-tubulin was the loading control. (E) Northern blot analysis of RNA from N2 and kgb-1 adults grown at 20°C or 26°C shows relative abundance of glh-4, glh-1 and kgb-1 transcripts, with an eIF-4A loading control. The kgb-1 probe did not include the 5' end of the gene, for which RT-PCR detects a small product in kgb-1(um3) RNA (not shown).