Fig. 4 ∣. Fasting induces Smpd3 expression to regulate cholesterol metabolism and VLDL production.
a, Hepatic mRNA expression of Smpd1, Smpd2, Smpd3 and Smpd4 in mice fasted for 4 h or 16 h (n = 6 and 7). b, Immunoblot analysis of hepatic SMPD3/nSMase2 in the livers of mice fasted for 4 h or 16 h (n = 3 and 3). Actin was used as a loading control. c, Immunoblot analysis of HA-tagged SMPD3/nSMase2 in the livers of mice transduced with AAV8-TBG-Ctr or AAV8-TBG-Smpd3 for 3 d (n = 3 and 3). Actin was used as a loading control. d,e, Hepatic SM (d) and CEs (e) in mice 3 d after i.v. injection of AAV8-TBG-Ctr or AAV8-TBG-Smpd3 (n = 5 and 4). f, Hepatic mRNA expression of indicated genes from WT mice transduced with AAV8-TBG-Ctr or AAV8-TBG-Smpd3 for 3 d (n = 5 and 5). g, VLDL-TG secretion in F/F control and L-A/C KO mice (n = 4 and 6). h. The rate of VLDL-TG production from F/F control and L-A/C KO mice (n = 4 and 6). i, Apo-B and ApoA-I protein levels in plasma of F/F and L-A/C KO mice (n = 4 and 4). j, TGs in isolated VLDL particles from mice treated with vehicle or Atglistatin (200 μmol kg−1) for 16 h (n = 5). All data are presented as mean ± s.e.m. P values were determined by two-sided Student’s t-test (d,e,h), two-sided Student’s t-test with Benjamini, Krieger and Yekutieli correction for multiple comparisons (a,f), or two-way ANOVA with Sidak’s correction for multiple comparisons (g,j). *P < 0.05, **P < 0.01, ***P < 0.001.