Journal: Journal of hazardous materials
Article Title: Mechanistic insight into airborne particulate matter PM10 as an environmental hazard for hemorrhagic stroke: Evidence from in vitro and in vivo studies.
doi: 10.1016/j.jhazmat.2024.136319
Figure Lengend Snippet: Fig. 5. PM10 promotes CA development through FasL-mediated necroptosis of VSMC in the cerebrovasculature. Representative immunofluorescence staining images of (a) p-RIPK1 and (b) FasL in 3D vascular spheroids following PM10 exposure. All sections were counterstained with anti-α −SMA antibody for VSMC as well as DAPI for nuclei (x400 magnification). Histograms represent the quantification of fluorescence intensity. The fluorescence intensity profile of FasL expression is shown. (c) Immunofluorescence staining for NF-κB p65 nuclear translocation in the vascular cell culture spheroid model system in response to PM10 (x400 magnification). The fluorescence intensity profile of the p65 localization is shown. (d and e) Assessment of the efficacy of the superoxide-dependent Fenton reaction in the vascular spheroids upon PM10 exposure, evaluated via fluorescent probes corresponding to intracellular labile iron pools (Calcein-AM) and superoxide radicals (HKSOX-1). (f) Immunohistochemical staining for p-MLKL in the internal carotid artery (ICA) (x400 magnification). (g) Immunofluorescence for p-RIPK3 with green fluorescence in ICA, counterstained with α-SMA (red) and DAPI (blue) (x400 magnification). (h) Immunofluorescence staining for NF-κB p65 nuclear translocation in the ICA following intratracheal PM10 instillation in mice (x400 magnification). The fluorescence intensity profile of the p65 localization is shown. (i) Representative immunofluorescence images for FasL upregulation in the ICA in response to PM10 exposure. (j) Serological levels of S100B, a diagnostic marker for hemorrhagic stroke, were assessed by ELISA in serum. (k) Immunofluorescence staining for myeloperoxidase (MPO) in the ICA, indicating increased circulating neutrophil infiltration into the cerebral vasculature following PM10 inhalation (x400 magnification). All data are presented as mean ± standard error of the mean, and *in dicates statistically significant differences between groups at *p < 0.05 (n = 6).
Article Snippet: HITB5 (CLU305), vascular smooth muscle cell (VSMC), was purchased from CEDARLANE (ON, Canada).
Techniques: Immunofluorescence, Staining, Fluorescence, Expressing, Translocation Assay, Cell Culture, Immunohistochemical staining, Diagnostic Assay, Marker, Enzyme-linked Immunosorbent Assay
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