MOESM1 of PEP-1-MsrA ameliorates inflammation and reduces atherosclerosis in apolipoprotein E deficient mice

Additional file 1: Figure S1. Vector diagrams of pET28a/MsrA and pET28a/PEP-1-MsrA. A 642-bp cDNA fragment of human MsrA was inserted into pET28a vector, PEP-1 was inserted at the N-terminus of MsrA

MOESM3 of PEP-1-MsrA ameliorates inflammation and reduces atherosclerosis in apolipoprotein E deficient mice

Additional file 3: Figure S3. Effects of MsrA or PEP-1-MsrA on body weight, spleen weight of apoE−/− mice. The 21-week-old apoE−/− mice were fed with a Western-type diet and intraperitoneally injected with PBS, MsrA or PEP-1-MsrA proteins for 12 weeks and then sacrificed. The body weights at various time-points (A) and spleen weights at the end point (B) were measured, n = 10

Bone marrow MCPIP1 deficiency dramatically reduced atherosclerotic lesion size in mouse aorta.

<p><b>A</b>. Representative atherosclerosis lesions of aortic root. <b>B</b>. Quantification of aortic root atherosclerotic lesion size. <b>C</b>. Representative <i>en face</i> aorta atherosclerosis.</p

Bone marrow MCPIP1 deficiency caused hematological disturbance.

<p><b>A</b>. Giemsa stained blood smears showed MCPIP1−/− bone marrow cell recipients contain dramatically reduced number of red blood cells. <b>B</b>. Vetscan HMT blood cell analysis showed reduced blood red cell (RBC) number, hemoglobin (HGB) concentration and hematocrit (HCT), as well as increased red cell distribution width (RDWc) which is calculated as Standard deviation ÷ mean cell volume x 100. <b>C</b>. Blood leukocyte profile showed increased number of neutrophils in MCPIP1−/− bone marrow cell recipients. <b>D.</b> Platelet related parameters of mice obtained by Vetscan HMT analysis. Parameters include total blood platelet count, platelet hematocrit (PCT), mean platelet volume (MVP), and platelet distribution width (PDWc) which is calculated using equation RDWc(%)  =  (Standard deviation ÷ mean cell volume) x 100. N = 6 (WT) or 9 (MCPIP1−/−).</p

MCPIP1 deficiency increased macrophage cholesterol efflux.

<p><b>A</b>. Bone marrow-derived macrophages were loaded with ac-LDL and cholesterol efflux to DMEM, apoAI and HDL was measured. Data were presented as mean ± SEM. N = 3 in each group. <b>B</b>. Bone marrow-derived macrophages were cultured in DMEM for 48 h with or without ac-LDL. Cell lysates were used for western blotting analysis for ABCA1 and ABCG1. Representative western blotting result was shown. <b>C</b>. Quantitative analysis of the western blots. Data were presented as mean ± SEM. N = 4 in each group. Open column: WT macrophages; Black column, MCPIP1−/− macrophages.</p

Bone marrow MCPIP1 deficiency increased mouse serum levels of proinflammatory cytokines and total and anti-oxLDL antibodies.

<p><b>A.</b> Serum levels of pro-inflammatory cytokines TNFα and IL-6 were determined by ELISA. <b>B.</b> Serum levels of total IgG and anti-oxLDL IgG were determined by ELISA as described in Materials and Methods.</p

Bone marrow MCPIP1 deficiency resulted in disorganization of mouse immune organs and leukocyte profile alteration in the spleen.

<p>Light microscopic images of mouse thymus, lymph nodes and spleen which were stained with hematoxylin and eosin. Magnification: 4X.</p

Bone marrow MCPIP1 deficiency caused multi-organ inflammation.

<p>Cryosections of kidney, liver and pancreas were stained with hematoxylin and eosin, and viewed with a Nikon Eclipse E600 microscope. Magnification: 40X</p

Bone marrow MCPIP1 deficiency altered leukocyte profile in immune organs.

<p><b>A and B.</b> Total number and flow cytometric analysis of immune cells in mouse thymus <b>(A)</b> and lymph nodes <b>(B).</b> *<i>p</i><0.05 vs WT group. <b>C.</b> Spleen weight and total spleen cell numbers of LDLR−/− mice received WT or MCPIP1−/− bone marrow cells. <b>D</b>. Leukocyte profile of splenocytes by flow cytometry. *p<0.01, n = 6 (WT) or 9 (MCPIP1−/−). <b>E.</b> Representative flow cytometric graphs of spleen CD11b+/Ly6C+ cells. Majority of the CD11b+/Ly6C+ cells are CD11b+/Ly6C^low (Blue box).</p

Bone marrow MCPIP1 deficiency resulted in growth retardation and lymphadenopathy.

<p><b>A</b>. Representative images of LDLR−/− mice received bone marrow cells from WT (WT) or MCPIP1 deficient (MCPIP1−/−) mice. <b>B</b>. Body weights of mice. *<i>p</i><0.01, n = 6 (WT) or 9 (MCPIP1−/−). <b>C</b>. Representative images of mouse lymph nodes.</p