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. 2014 Dec 15;34(3):344–360. doi: 10.15252/embj.201490464

Figure 3.

Figure 3

miR-379 controls systemic triglyceride levels via the lipolysis-stimulated lipoprotein receptor (LSR) and the low-density lipoprotein receptor (LDLR)
  1. Serum 3H-triolein in a lipid tolerance test in db/db mice treated with an anti-miR-379 or scrambled control Tough Decoy (TuD) construct delivered by rAAV (n = 5–6). Mice were fasted for 16 h and given a 100-μl oral fat load of olive oil spiked-in with 3H-triolein. Inset: area under the curve. Line graphs show mean ± SEM; ANOVA (with post hoc test): ***P < 0.001 or **P < 0.01.
  2. Organ distribution of 3H-triolein radioactivity from animals in (A). BAT, brown adipose tissue; pgWAT, perigonadal white adipose tissue; GCM, gastrocnemius skeletal muscle. Bar graphs show mean ± SEM; t-test: *P < 0.05.
  3. Protein levels of LSR and LDLR from livers of animals treated with anti-miR-379 or scrambled control LNA (n = 5), same animals as in Fig2A. Shown are the LSR-α (68 kDa) and LSR-β (56 kDa) subunits and the glycosylated LDLR (between 100 and 130 kDa) protein.
  4. Protein levels of LSR and LDLR from livers of wild-type (wt) mice treated with dexamethasone (1 mg/kg BW) or isotonic saline for 28 days. (n = 7–8).
  5. Western blot of liver extracts from wt or LDLRKO mice treated with control or LSR shRNA-containing adenovirus and with anti-miR-379 or scrambled control LNA (n = 7).
  6. Serum triglyceride levels of same animals as in (E) under fed conditions. Bar graphs show mean ± SEM; ANOVA (with post hoc test): **P < 0.01 or *P < 0.05.
  7. Flag-LSR protein levels in HEK293 cells treated with miR-379 mimics. 100 nM cel-miR-293b was used as the control mimic. [-]: untransfected.
  8. miR-379 target validation of LSR (H) and LDLR (I) using a dual-luciferase reporter gene assay. miRNA binding site (MBS) predicted by RNA22 and MiRTiF and the corresponding mutated (mut) sequence were cloned into psiCHECK™-2 vector. Bar graphs show mean ± SEM; ANOVA (with post hoc test): ***P < 0.001.