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The other half was fixed with 10% Formalin (Sigma) for 16 h at 4 C, transferred to a 70% v/v ethanol solution for a further 24 h and was then paraffin-embedded for immunohistochemistry

As fat decreases, the firm gland often becomes more noticeable

The mutant strains unc-16 ( e109 ), exp-1 ( sa6 ), and egl-8 ( sa47 ) were obtained from the Caenorhabditis Genetics Center (University of Minnesota, Minneapolis, MI, USA), and had been used without further outcrossing to the wild-type strain