Extended Data Fig. 2 ∣. Fasting stimulates hepatic PM–ER cholesterol transport.

a, Gross appearance of livers from mice fasted for 4- or 16-h. b, Hepatic triglycerides in mice fasted for 4- or 16-h (n = 3/3). c, Hepatic CE in mice fasted for 4- or 16-h (n = 3/3). d, Significantly upregulated pathways in the livers of mice fasted for 16-h compared to 4-h according to pathway analysis of RNA Sequencing data. e, Significantly downregulated pathways in the livers of mice fasted for 16-h compared to 4-h according to pathway analysis of RNA Sequencing data. f, Hepatic mRNA expression of SREBP-2 pathway targets from the livers of mice fasted for 4- or 16-h (n = 3/3). g, Quality control of plasma membrane isolation from the mouse liver. Cadherin: PM maker; Calnexin: ER maker; ATGL: lipid droplet (LD) maker; Actin: cytoskeleton maker. This analysis was completed once as the organelle isolation method has been previously validated (further method details are in the Methods section). h, Free cholesterol analysis from purified PMs of wild-type mice (n = 3/3). i, TLC analysis of free cholesterol (FC), sphingomyelin (SM) and phosphatidylcholine (PC) from the livers of mice fasted for either 4 or 16 h. j, PM total lipids as measured by mass-spec from livers of F/F control and L-A/C KO after 4- and 16-h fasting (n = 3/3). All data are presented as mean ± SEM. P values were determined by two-sided Student’s t-test (b, c and h), or two-sided Student’s t-test with Benjamini, Krieger and Yekutieli correction (f).