Extended Data Fig. 9: Workflow diagram of fecal microbiota transplantation from human donors to germ-free mice littermates.

a, Workflow for the preparation of fecal microbiota slurry. Stool (250 mg) from both anorexia (AN) and control (HC) was cut on dry ice, transferred to anaerobic chamber, and resuspended with 5 ml of LYBHI media diluted in 20% glycerol. The resuspended fecal slurries were aliquoted in cryotubes and refrozen back quickly at -80 °C until further use. All stool samples of matched AN and HC donors were prepared on the same day and frozen aliquots were stored frozen for gavage to mice. b, Experimental scheme for the GF mice transplantation study. In each independent litter study, GF female littermates at age of six weeks old were taken out of breeding isolator and were randomly assigned to receive 200 µl of fecal slurries from three AN cases or three HC subjects. Both groups of mice were housed in autoclaved individually ventilated cages and were given autoclaved chow diet and water ad libitum for two days. After two days, mice were gavaged with a second dose of fecal material from the same matched AN and HC donors as before. Thereafter, mice in both groups were single housed and subjected to 30% calorie restricted chow diet for three weeks. Water was given ad libitum during this period. Both the anorexia-transplanted (AN-T) and the normal control-transplanted (HC-T) mice were weighed every five days after the start of energy-restricted diet. Created with Biorender.com.