Fig. 5. CeMM culture results in decreased lipid and protein stores. (A) Nile Red staining of lipid deposits in the intestinal cells reveals decreased lipid stores. Almost all animals display strong staining when fed on NGM and decreased staining when starved for 24 h or fed CeMM. All images had the same exposure time. The percentage of 100 animals per condition displaying similar fluorescence intensity is shown on the right of each image. Images of animals fed CeMM are only of the gut immediately posterior to the pharynx. This is the most fluorescent portion of the gut as seen in the image of the NGM fed animal. The CeMM fed group fall into three categories: normal fluorescence (top), weak fluorescence (middle) and very weak fluorescence (bottom). Assignment to these three groups reflects the relative exposure time (RE) needed to produce an image with normal fluorescence top (RE=1), middle (RE=2) bottom (RE=4). Starved NGM have an RE=2 whereas starved CeMM have an RE=4. Note the fed NGM image is saturated at RE=1. (B) Staining for ß-galactosidase activity in unc-54::lacZ transgenic animals reveals decreased protein stores. The percentage of 100 animals per condition displaying the staining pattern is shown on the right of each image. Almost all animals display full body wall muscle staining when fed on NGM and no staining when starved for 48 h (stain near the vulva is embryo staining). Most animals in CeMM display no body wall muscle staining but head and vulval muscle staining, as associated with 24 h starvation (Zdinak et al., 1997). A smaller percentage of animals display body wall muscle staining when fed CeMM and an even smaller percentage no staining. In contrast to the degradation of this reporter seen in starved animals, microarray analysis suggests the decreased staining is due to decreased synthesis alone.