Additional file 1 of Bilirubin ameliorates murine atherosclerosis through inhibiting cholesterol synthesis and reshaping the immune system
Creators: name: Wen, Guanmei
name: Yao, Leyi
name: Hao, Yali
name: Wang, Jinheng
name: Liu, Jinbao
Publisher: name: figshare
Year: 2022
Subjects: subject: Medicine
subject: Genetics
subject: FOS: Biological sciences
subject: Physiology
subject: 39999 Chemical Sciences not elsewhere classified
subject: FOS: Chemical sciences
subject: Immunology
subject: FOS: Clinical medicine
subject: 69999 Biological Sciences not elsewhere classified
subject: Science Policy
subject: 110309 Infectious Diseases
subject: FOS: Health sciences

description: Additional file 1: Figure S1. (A) ApoE−/− mice fed with a western-type diet were intraperitoneally injected with or without bilirubin. After 10 weeks, mice were scarified, and the body and liver weight were measured. The relative liver weight was expressed as liver weight to body weight ratio. (B) Concentrations of liver enzymes were determined by biochemical analysis. Error bars represent mean ± SD. n (control) =8, n (bilirubin) =14. *P &lt; 0.05. Differences were determined by an unpaired t-test. Figure S2. (A) Quantification of hepatic HMGCR protein expression in ApoE−/− mice treated with bilirubin (n=3) or vehicle (control, n=3). (B) BALB/c mice were received intraperitoneal injection of bilirubin (20 mg/Kg/day, n=4) or vehicle (n=4) once a day for 3 days and sacrificed at day 4. The expression levels of HMGCR in the livers were determined by western blot. Bar plots showing the relative HMGCR protein expression in the livers (right panel). *P &lt; 0.05. Figure S3. After determining the expression of 27 surface markers on the spleen cells using mass cytometry, the T cells expressing CD3 were gated and analyzed using viSNE. viSNE was used to visualize the distribution of the spleen T cells from both groups. Cells on the viSNE map were colored by normalized expression of indicated surface markers. (B) Bar plots showing the frequencies of T1 or T2 in spleen T cells obtained from ApoE−/− mice treated with or without bilirubin. Error bars represent mean ± SD. n (control) =8, n (bilirubin) =14. Figure S4. (A) Dot plots showing the Pearson correlation coefficients for relationships between the concentrations of LDL or TCHO in peripheral blood and frequencies of PMN-MDSCs or All MDSCs. (B) Dot plots showing the Pearson correlation coefficients for relationships between the concentrations of LDL in peripheral blood and the frequencies of Ly6C+NK cells. (C) Dot plots (n=22) showing the Pearson correlation coefficients for relationships between the concentrations of LDL or TCHO in peripheral blood and the frequencies of peripheral blood B cells. Correlations were determined by a Pearson test. Figure S5. After determining the expression of 27 surface markers on the PWB cells using mass cytometry, the T cells expressing CD3 were gated and analyzed using viSNE. (A) viSNE was used to visualize the distribution of the peripheral blood T cells from both groups. Cells on the viSNE map were colored by normalized expression of indicated surface markers. (B) Bar plots showing the frequencies of T1 or T2 in peripheral blood T cells obtained from ApoE−/− mice treated with or without bilirubin. (C) Bar plots showing the frequencies of T28 or T29 in peripheral blood T cells obtained from ApoE−/− mice treated with or without bilirubin (left panel). Dot plots showing the Pearson correlation coefficients for relationships between the concentrations of total bilirubin in peripheral blood and frequencies of T28 or T29 in peripheral blood T cells. (D) Dot plots (n=22) showing the Pearson correlation coefficients for relationships between the concentrations of TCHO in peripheral blood and the frequencies of T7, T9, T13 in peripheral blood T cells. Error bars represent mean ± SD. n (control) =8, n (bilirubin) =14. Differences were determined by an unpaired t-test. Correlations were determined by a Pearson test. Table S1. Mass cytometry antibody reagents.