Extended Data Fig. 7: nt-LNA treatment promotes atherosclerotic regression in Ldlr-/- mice.
From: LDL delivery of microbial small RNAs drives atherosclerosis through macrophage TLR8
(a) Schematic for regression study design. Male(M) and female(F) Ldlr-/- mice were fed a chow diet (n = 6 mice) or an atherogenic diet (n = 50 mice) for 14 weeks. After 14 weeks, chow-fed mice and a subset of mice from the atherogenic diet group (baseline; n = 7 M/8 F mice). Remaining diet-fed mice were then switched to a chow diet to allow lesion regression (Reg.) and were injected once weekly with saline control (Reg. Ctr; n = 9 M/9 F mice) or Reg. nt-LNA (30 mg/kg; n = 9 M/8 F mice). b) Plasma total cholesterol (TC) or c) triglycerides (TG) following fractionation by SEC; chow (n = 6; 3 M/3 F), baseline (n = 8; 4 M/4 F), Reg. Ctr; (n = 10; 5 M/5 F) and Reg. nt-LNA (n = 10; 5 M/5 F) d) Immunoblots of plasma proteins in Reg. Ctr (n = 10) or Reg. nt-LNA (n = 9) groups. Representative images of two independent experiments are shown. e) Quantification of immunoblots by densitometry. f-g) Lesion area of serial sections of the aortic root in baseline (n = 15 mice; 7 M/8 F), Reg. Ctr(n = 18 mice; 9 M/9 F) or nt-LNA (n = 17; 9 M/8 F) groups. h) Lesion area under the curve (AUC) for both sexes of mice as determined by Oil Red O staining in the aortic root (Baseline: n = 15; Reg. Ctr: n = 18; Reg. nt-LNA; n = 17). One-way ANOVA; Dunnett’s multiple comparison test, **p < 0.01, ***p < 0.001. i) Lesion AUC for mice of each group separated by sex. Two-way ANOVA; Dunnett’s multiple comparison test, **p < 0.01. j-k) Masson’s Trichrome staining and quantification of fibrosis in aortic roots of baseline (n = 7 mice; 3 M/4 F) Reg. Ctr (n = 9 mice; 4 M/5 F) or Reg. nt-LNA (n = 10 mice; 5 M/5 F) groups. l-m) MAC2 (green) immunofluorescence and quantification within aortic roots obtained of at baseline (n = 7 mice; 3 M/4 F), Reg. Ctr (n = 10 mice; 5 M/5 F), or Reg. nt-LNA (n = 10 mice; 5 M/5 F) groups. Two-way ANOVA; Tukey’s multiple comparison test, **p < 0.01, ***p < 0.001. Data are mean ± SEM. Scale bar = 500 μm. Numerical source data, statistics, exact p values and q values are provided.
