Journal: Journal of Tissue Engineering

Article Title: Formation of neutrophil extracellular traps in the early stages exacerbate the healing process by regulating macrophage polarization in Achilles tendon-bone injury

doi: 10.1177/20417314251348038

Figure Lengend Snippet: PAD4 inhibition reduces M1 macrophage polarization after tendon-bone injury. (a) Immunohistochemistry (IHC) analysis of CD68 in tendon-bone section from control, TI, and TI + PAD4 inhibitor groups. (b) Quantification of CD11c positive cells. n = 6 mice/group. (c) IHC analysis of CD11c in tendon section from control, TI, and TI + PAD4 inhibitor groups. (d) Quantitative analysis of CD11c positive cells in c. n = 6 mice/group. (e) Western blot of iNOS, TLR4 and IL-1β in tendon tissue from control, TI, and TI + PAD4 inhibitor groups. (f) Quantification of relative gray intensity/GAPDH in c. n = 3 mice/group. Data are presented as the mean ± SEM. Scale bar = 1 mm. Significance was examined by one-way ANOVA (b, d, and f). ** P < 0.01, **** p < 0.0001.

Article Snippet: For M1 macrophages detection, sections were incubated with CD68 (14-0681-82; Invitrogen), CD11c (14-0114-82, Invitrogen), and iNOS (MAB9502; R&D systems) antibodies.

Techniques: Inhibition, Immunohistochemistry, Control, Western Blot

Journal: Cell Transplantation

Article Title: Intravenous Administration of Human Amniotic Mesenchymal Stem Cells in the Subacute Phase of Cerebral Infarction in a Mouse Model Ameliorates Neurological Disturbance by Suppressing Blood Brain Barrier Disruption and Apoptosis via Immunomodulation

doi: 10.1177/09636897211024183

Figure Lengend Snippet: The administration of hAMSCs suppresses pro-inflammatory immune cells around the peri-infarct area. (a) Tissue section stained with 4′,6-diamino-2-phenylindole (DAPI) showing regions of analysis in the peri-infarct area ((a) ischemic core). Three random regions, indicated by solid boxes (200× magnification), per coronal section and two coronal sections per brain were analyzed to count iNOS+/CD11bc+ cells [M-high: n = 3; M-vehi: n = 3]. Scale bar: 200 µm. (b) Representative images of the immunofluorescent staining of brain sections in the M-high and M-vehi mice. iNOS (green) was colocalized (yellow) with CD11b/c (red). Nuclei were counterstained using DAPI (blue). Scale bar: 50 µm. 200× magnification. (c) Cell count of iNOS positive, CD11b/c positive, and iNOS/CD11bc colocalized cells. Scale bar: 50 µm. ** P < .01, *** P < .001, unpaired t -test.

Article Snippet: The sections were incubated overnight at 4°C with an anti-iNOS antibody (1:500; Funakoshi, Tokyo, Japan, MAB9502), or an anti-CD11b/c antibody (1:100; Bioss, Woburn, USA, bs-1014R), and then incubated with corresponding Alexa Fluor 488- or 555-conjugated secondary antibodies (Cell Signaling Technologies, Danvers, Massachusetts, USA) at room temperature for 2 h. Nuclei were counterstained with 4′,6-diamino-2-phenylindole (DAPI; Sera Care, Milford, Iowa, USA).

Techniques: Staining, Cell Counting