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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).
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