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. 2021 Oct 8;6(19):e147038. doi: 10.1172/jci.insight.147038

Figure 2. VLDL/LDL secretion is reduced accompanied with less microsomal triglyceride transfer protein in ASGR1-deficient mice.

Figure 2

Hepatic lipid secretion assays were conducted on mice with different genotypes. (A) TG and (B) cholesterol content were measured at 0, 1, 2, and 3 hours after i.p. injection with tyloxapol, n = 5. (C) The relative mRNA expression level of secretion-related genes, Apob and microsomal triglyceride transfer protein (Mttp) were analyzed by real-time PCR (RT-PCR) in liver tissues (WT, n = 7; Asgr1+/–, n = 4; Asgr1–/–, n = 7). (D) The corresponding proteins, APOB and MTTP, were analyzed by Western blots in liver tissues. Representative images of the immunoblotting and gray intensity of each band relative to β-actin was shown, n = 3. (E) Secreted APOB content in mouse serum was analyzed by Western blots (WT, n = 3; Asgr1+/–, n = 4; Asgr1–/–, n = 4). Parallel analyses were conducted on HepG2 cell lines. (F) The mRNA expression level of APOB and MTTP was analyzed by RT-PCR (n = 4), and (G) protein content of APOB and MTTP was analyzed by Western blots, n = 4. (H) ELISA comparison of secreted APOB content between WT and ASGR1–/–, n = 4. All data are shown as the means ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 as compared with the indicated WT by 1-way ANOVA among 3 groups. Statistical significance between 2 groups was assessed with an unpaired, 2-tailed t test.