Fig 1: In vivo immune response of scaffold in mouse intramuscular and rabbit skull defect model.a H&E staining of implants on days 7 and 14 after implantation in mouse intramuscular. b Cell morphologies of macrophages on different implants. c1) Representative flow cytometry plots of F4/80+, CD197+CD206- macrophages (M1) and CD197-CD206+ macrophages (M2) polarization in DCLS and HCLS on days 7 and 14 after implantation in mouse intramuscular. c2–4) Relevant quantification of F4/80+, CD197+CD206- macrophages (M1) and CD197-CD206+ macrophages (M2) polarization in DCLS and HCLS (c2: **p = 0.0054, ***p = 0.0004, ***p = 0.0009, *p = 0.0431; c3: **p = 0.0078, ***p = 2.1538 × 10-5, ***p = 0.0006, ns = 0.1301; c4: **p = 3.7781 × 10-5, ***p = 6.6342 × 10-5, *p = 0.0482, *p = 0.0365). d1–5 Expression of inflammatory factors induced by implants after implantation in the mouse intramuscular model for 7 and 14 days. n = 3 biologically independent replicates (d1: *p = 0.0378, **p = 0.0018, **p = 0.0022; d2: **p = 0.0065, ***p = 7.8921 × 10-5, ns = 0.0843; d3: **p = 0.0024, **p = 0.0019, ***p = 2.4562 × 10-5; d4: *p = 0.0342, **p = 0.0055, ***p = 4.1243 × 10-7; d5: *p = 0.0158, *p = 0.0301, **p = 0.0087). e1 Representative immunostainings of CD197 and CD206, BMP-2 secretion and F4/80+, and VEGF secretion and F4/80+ macrophages on days 7 and 14 after implantation in mouse intramuscular. e2–5 Relevant semiquantitative analysis of immunofluorescence staining by Image J software e2: *p = 0.0180, **p = 0.0091; e3: *p = 0.0342, *p = 0.0435; e4: **p = 0.0033, ***p = 8.0122 × 10-6; e5: *p = 0.0185, **p = 0.0044). f1 Representative immunostainings of CD197 and CD206, BMP-2 secretion and F4/80+ and VEGF secretion and F4/80+ macrophages on days 7 and 14 after implantation in rabbit skull defect. f2–5 Relevant semiquantitative analysis of immunofluorescence staining by Image J software (f2: **p = 0.0020, *p = 0.0416; f3: *p = 0.0196, *p = 0.0394; f4: *p = 0.0149, ***p = 5.5420 × 10-8; f5: ***p = 4.6347 × 10-6, ***p = 8.1728 × 10-7). n = 3 cells examined three independent experiments. (Two-sided comparison, error bars represent standard deviation, *p < 0.05, **p < 0.01, and ***p < 0.001).
Fig 2: Representative Flow Cytometry Plots Comparing Endogenous and Transferred Myeloid Cells in TumorsCells were pregated on live singlets and identified as either F4/80+ tdT- (endogenous) or F4/80+ tdT+ (transferred) tumor-associated macrophages (TAM). The expression of CD11c and MHCII within each TAM population is compared in the final plot.
Fig 3: Expression of Shh and macrophage infiltration in control and LysMCre/SmoKO mice.Immunohistochemistry of Shh expression in A control and B LysMCre/SmoKO mouse stomachs. Shh expression at the ulcer margin of C, D control, and E, F LysMCre/SmoKO mouse stomachs. G Shh concentrations (pg/mL) measured in plasma collected from in control and LysMCre/SmoKO mice. Macrophage numbers (CD11b+F4/80+Ly6Chi) within the uninjured and injured gastric epithelium of H control and I LysMCre/SmoKO mice 1–7 post-ulcer induction. *P < 0.05 compared to day 1 post-injury, n = 6 mice per group. Data are shown as the mean ± SEM.
Fig 4: Tick EVs bind to mammalian immune cells.a Overrepresentation of proteins involved in cell adhesion signaling networks in I. scapularis salivary gland EVs. The biological relationship between proteins was determined using the right-tailed Fisher’s exact test with Benjamini–Hochberg multiple-testing correction. p < 0.05; -log (p value) >1.5. b Graphic representation of focal adhesion and integrin signaling proteins (purple) found in I. scapularis salivary gland EVs. Confocal images of c murine bone marrow-derived macrophages (BMDMs) (bar = 20 µm) and g macrophages derived from human peripheral blood mononuclear cells (green) bound to I. scapularis EVs labeled with PKH26 (orange) (bar = 20 µm), respectively. Cells were pre-treated with cytochalasin D (5 µM). (-) No EVs; (+) EVs. Images are representative of two independent experiments. d Arbitrary fluorescent units (AFU) of murine BMDMs bound to PKH26-labeled tick EVs (red). BMDMs incubated in the absence of EVs (black). Each time point represents the mean fluorescence from seven different cells minus background and normalized to time 0. Graphs show one of two independent experiments. Flow cytometry with e murine BMDMs (F4/80+-APC) and h human macrophages (CD11b+-APC/Cy7) bound to DiO-labeled tick EVs (40 µg), respectively. No EVs (-); EVs (+). Figures are representative of three biological replicates. f Flow cytometry analysis of unlabeled BMDMs (black), F4/80-APC-labeled BMDMs (blue), F4/80-APC-labeled BMDMs bound to 40 µg DiO-labeled tick EVs (red), and F4/80-APC-labeled BMDMs bound to 20 µg of DiO-labeled and 20 µg unlabeled tick EVs (gray). The histogram shows one of three biological replicates. PKH26 is a red fluorescent dye with long aliphatic tails45. In c, g, PKH26 was artificially transformed to an orange color to be visualized by color-blind readers. Source data are provided as a Source Data file.
Fig 5: Microglial-derived MP are increased in the blood following TBI. Flow cytometry analysis of enriched MP in the blood from sham and TBI mice at 24 h post-injury. a Representation of gating strategy used to characterize MP using SSC-H and standard microbeads (300- to 1000-nm diameter). Standard microbeads (P1 gated population) were used as an internal control to determine the size of MP in the blood, and annexin V staining confirmed MP characteristics. At 24 h post-injury, total blood MP is increased in TBI mice compared with sham-injured mice. b Measurements of leukocyte-derived (CD18), macrophage-derived (F4/80), and microglial-derived (P2Y12/CD45) MP in the blood from sham and TBI mice at 24 h post-injury. Microglial-derived MP are significantly increased in TBI mice when compared with sham-injured mice (*p < 0.5 vs sham; Student’s t test; n = 6/group). Bars represent mean ± standard error of the mean (S.E.M.). Data represent results of three independent experiments
Supplier Page from Thermo Fisher Scientific for F4/80 Antibody APC