Table 2.

Mouse characteristics

	Group				ANOVA P Values
	SC-C	SC-DMB	WD-C	WD-DMB	Interaction	Diet	Treatment
n	13	13	11	13
Water intake, mL/day	3.8 ± 0.1	3.5 ± 0.2	2.6 ± 0.1*	2.0 ± 0.1†	0.39	<0.0001	0.01
Plasma concentrations of TMAO-related metabolites
Choline, µM	36 ± 3	39 ± 4	93 ± 9*	74 ± 6b	0.07	<0.0001	0.17
l-carnitine, µM	23 ± 2	18 ± 2	11 ± 1*	9 ± 2	0.65	<0.0001	0.06
Betaine, µM	86 ± 21	68 ± 14	46 ± 9a	28 ± 5	0.98	<0.01	0.19
Body mass at euthanasia, g	30.9 ± 0.9	30.4 ± 0.7	40.7 ± 2.0*	36.8 ± 1.3b	0.18	<0.0001	0.09
Mass of key organs
Total heart, mg	133 ± 3	132 ± 3	156 ± 4*	152 ± 6	0.69	<0.0001	0.58
Left ventricle, mg	86 ± 4	88 ± 2	96 ± 3*	97 ± 4	0.81	<0.01	0.61
Lungs, mg	165 ± 5	163 ± 5	191 ± 6*	189 ± 6	0.97	<0.0001	0.69
Liver, g	1.5 ± 0.1	1.5 ± 0.03	2.7 ± 0.3*	2.2 ± 0.2†	0.11	<0.0001	0.08
Kidneys, mg	408 ± 9	417 ± 19	449 ± 14a	454 ± 8	0.87	<0.01	0.59
Visceral fat, g	0.9 ± 0.1	0.7 ± 0.1	2.4 ± 0.2*	2.0 ± 0.2	0.42	<0.0001	0.08
Spleen, mg	72 ± 2	74 ± 4	105 ± 5*	91 ± 2†	0.02	<0.0001	0.12
Brain, mg	397 ± 13	402 ± 12	447 ± 9*	443 ± 9	0.70	<0.001	0.95
Skeletal muscle mass
Quadriceps, mg	339 ± 17	349 ± 20	416 ± 11*	422 ± 6	0.89	<0.0001	0.60
Gastrocnemius, mg	270 ± 7	269 ± 9	328 ± 6*	321 ± 4	0.59	<0.0001	0.60
Tibialis anterior, mg	101 ± 6	102 ± 6	74 ± 5*	74 ± 6	0.94	<0.0001	0.91
Soleus, mg	19 ± 1	18 ± 2	21 ± 1	23 ± 1	0.22	0.04	0.57
Food intake, kcal/day	12.5 ± 0.3	12.6 ± 0.2	13.6 ± 0.3*	12.5 ± 0.2†	<0.01	0.04	0.05
Norm. food intake, kcal/day/g BW	0.43 ± 0.01	0.42 ± 0.01	0.36 ± 0.01*	0.36 ± 0.01*	0.45	<0.0001	0.48
Artery diameters, µm
Carotid artery resting	447 ± 9	435 ± 6	436 ± 10	430 ± 8	0.73	0.30	0.36
Carotid artery, maximal	453 ± 9	443 ± 7	455 ± 11	448 ± 8	0.86	0.36	0.67
Aorta, resting	646 ± 15	604 ± 13	627 ± 19	608 ± 21	0.53	0.67	0.09

Data are mean ± SE. Groups are standard chow-control water (SC-C), standard chow-DMB water (SC-DMB), Western diet-control water (WD-C), and Western diet-DMB water (WD-DMB). Statistics are two-way ANOVA with Šidák’s multiple comparisons tests. Symbols denote pairwise comparisons as follows: *P < 0.05 vs. SC-C; †P < 0.05 vs. WD-C; ^aP < 0.10 vs. SC-C; ^bP < 0.15 vs. WD-C. Statistical outliers (ROUT: Q = 1%) were removed as follows: two for l-carnitine (1 SC-C group, 1 SC-DMB); two for kidney and spleen masses—these data were from the same two mice in the WD-DMB group who had enlarged kidneys and spleens (the WD-DMB group average with these mice still included were 473 ± 15 mg for kidneys and 112 ± 15 mg for spleen); and one from aorta diameter (SC-DMB). BW, body weight; TMAO, trimethylamine N-oxide.