Extended Data Fig. 1: Aspa is highly expressed in differentiated adipocytes and across adipose tissue depots.
From: N-acetylaspartate from fat cells regulates postprandial body temperature
a, Aspa gene expression across mouse tissues (n = 3 mice/tissue). Data represent mean ± s.e.m. *P < 0.05, **P < 0.01, ****P < 0.0001 by ordinary one-way ANOVA followed by Dunnett’s multiple comparisons test of brain vs different tissues. Expression of Aspa and adipogenic genes (Adipoq, Fabp4, and Pparg2) in pre-adipocytes (Pread) and mature adipocytes (Adipo) in mice: (b) SVF-derived adipocytes, (c) 3T3-L1 adipocytes (n = 3 biological replicates/group). Data represent mean ± s.e.m. d, Expression of ASPA and adipogenic genes (ADIPOQ, FABP4, and PPARG2) in human primary pre-adipocytes (Pread) and mature adipocytes (Adipo) (n = 3 biological replicates/group). Data represent mean ± s.e.m. e, Relative abundance of NAA and TCA metabolites in 3T3-L1 pre-adipocytes (Pread) and mature adipocytes (Adipo) (n = 5 biological replicates/group). Data represented as box-and-whisker plots using the Min-to-Max method in GraphPad Prism: box limits, 25th to 75th percentiles; center line, median; whiskers, minimum and maximum values. f, Immunoblot of 3T3-L1 cells and human primary cells probed for ASPA, ADIPOQ, and PPARγ before (-d) and after adipocyte differentiation (+d). HSP90 served as the loading control. g, Immunofluorescent staining of ASPA (red) and co-labeling of lipid droplets (green, LipidTox), and nuclei (blue, DAPI) of primary human adipocytes before and after adipocyte differentiation. h, Correlation between expression of ADIPOQ and ASPA in WAT of people before and after weight loss (WL) (n = 15 samples/group). P-value for Pearson’s r (r) <0.001 by one-sided t-test performed on the linear regression. b-e, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 by unpaired two-tailed Student’s t-test. f,g representative of 3 independent experiments.
