Researchers reveal why fat increases after dieting and how protein might help
, confirmed by sequencing of full-length 16S rRNA genes and stored in PBS containing 6% glycerol at −80 °C until further use. Germ-free or gnotobiotic mice at age of 9 weeks were gavaged with 0.2 ml LC2W or Lam-1 for three consecutive days and then fed with 10%, 25% and 65% food in the next 3 d. After feeding ad libitum for 24 h, mice were gavaged with BODIPY-labeled fatty acid and feces, blood and the tissues of interest were collected at the indicated time points.
Metabolites in serum with variable importance in the projection values >1 obtained from the orthogonal partial least-squares discriminant analysis model andvalues 0.1% and LDA score >2 between two groups) were performed in GraphPad Prism to generate heat maps.Lactobacillus LC2W and Lam-1 cultured in MRS medium in the early stationary growth phase with a concentration of approximately 10c.f.u. per ml were collected. After centrifugation of 2 ml bacterial cultures, the resultant bacterial pellets were mixed with 400 µl HO, 800 µl methanol and 1.2 μl formic acid. A volume of 0.4 ml cell-free supernatant or 0.4 ml LB or MRS medium was mixed with 800 µl methanol and 1.2 μl formic acid.
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