Fig. 2
From: Methyl group donors abrogate adaptive responses to dietary restriction in C. elegans
Choline and methionine supplementation do not abrogate the enlargement of the hypodermal lipid droplets of dietarily restricted worms. a Hypodermal lipid droplets in the heads of ad libitum-fed L4 larvae (a, c, e) and dietarily restricted L4 larvae (b, d, f) without (H2O vehicle control; a, b) or with choline (c, d) and methionine (e, f) supplementation were visualized using short-term vital BODIPY™ 493/503 staining. Image stacks with a depth of 10 μm, consisting of 21 focal planes at 0.5-μm intervals, were captured and rendered as maximum projections. Yellow arrowheads indicate the beginning of the intestine. The BODIPY™ 493/503 fluorescence signals (green) and the auto-fluorescence signals of lysosome-related organelles (red) are shown to study possible co-localizations of the signals. Scale bars, 20 and 10 μm (magnifications). b–f Quantification of hypodermal lipid droplets in the head using the 3D spot segmentation plug-in of ImageJ software. The mean LD volume (b), the summarized LD number/area (AU, arbitrary unit) (c), and the summarized LD volume/area (AU, arbitrary unit), a proxy indicator of the extent of fat storage (D), are shown. The distributions of the relative LD number e and LD volume f per LD size category (<1, 1–3, 3–5, >5 μm3) are shown. The data were derived from six to nine individual worms per feeding condition in three independent experiments. AL ad libitum, AL CH ad libitum condition with choline supplementation, DR dietary restriction, DR CH dietary restriction condition with choline supplementation