mouse endothelial antigen meca 32  (Developmental Studies Hybridoma Bank)

Journal: Physiological Reports

Article Title: Blocking P2X7 receptor with AZ 10606120 exacerbates vascular hyperpermeability and inflammation in murine polymicrobial sepsis

doi: 10.14814/phy2.15290

Figure Lengend Snippet: Blocking P2X7 receptor with AZ 10606120 does not prevent lung apoptosis during murine sepsis. Mice were subjected to cecal slurry (CS) to induce sepsis in the presence or absence of 1‐h pre‐treatment with AZ 10606120 (AZ; 10 µg/g, SQ) for 24 h. (a) Representative images of apoptotic (TUNEL) fluorescence staining are shown at 10X and 60X (insert) power [blue = DAPI (nuclei), red = MECA‐32 (endothelial cells), green = TUNEL (apoptotic cells)]. (b) Quantification of TUNEL‐positive fluorescence staining shows AZ 10606120 did not prevent apoptosis in lung tissue of mice subjected to CS. There were no statistical differences in the (c) number or (d) percentage of TUNEL‐positive cells that were endothelial (overlap of MECA‐32 [red] on TUNEL [green]). Statistical analysis was performed using one‐way ANOVA with Tukey's multiple comparisons. Graphs represent mean ± SEM ( n = 4–8)

Article Snippet: Slides were incubated in the dark with TUNEL reaction mixture at 37°C for 1 h. Slides were rinsed three times with PBS, blocked with 10% normal goat serum in PBS and 0.05% Tween‐20 for 1 h, and incubated with rat anti‐mouse monoclonal MECA‐32 (panendothelial) antibody (1:100, BioxCell BE0200) in PBS, 3% normal goat serum, 0.05% Tween‐20 overnight at 4°C for endothelial labeling.

Techniques: Blocking Assay, TUNEL Assay, Fluorescence, Staining