Two experiments were conducted to determine the effect of dietary fiber source and level on egg yolk, liver, and plasma cholesterol concentrations of White Leghorn laying hens. Initially, dietary fiber levels of 2.05, 4.41, 6.68, and 8.79% furnished mainly by sunflower meal were fed to laying hens for 140 days. In the second experiment, alfalfa meal, ground whole oats, sunflower meal, rice mill feed, or wood shavings was added to a corn-soybean meal basal diet to furnish 2.00% added crude fiber and fed to laying hens for 84 days. Yolk cholesterol decreased 4.39, 10.38, and 13.29% by feeding crude dietary fiber levels of 4.41, 6.68, and 8.79%, respectively, to hens as compared to a corn-soybean meal basal diet containing 2.05% crude fiber. Egg yolk cholesterol was significantly decreased by feeding alfalfa meal, oats, sunflower meal, rice mill feed, or wood shavings to laying hens when compared to yolk cholesterol of hens fed the basal diet. The greatest reduction in egg yolk cholesterol was found by feeding either oats or wood shavings. No significant differences were found in plasma cholesterol due to dietary fiber level. Plasma triglycerides decreased and liver cholesterol increased as dietary fiber level increased in diets fed to laying hens. When laying hens were fed alfalfa meal, oats, rice mill feed, or wood shavings, plasma cholesterol significantly decreased. Liver cholesterol increased when hens were fed either alfalfa meal or rice mill feed as the primary fiber source.