Journal: International Journal of Molecular Sciences

Article Title: Generation of Functionally Competent Human Mast Cells from Limited Blood Volumes

doi: 10.3390/ijms27041793

Figure Lengend Snippet: PB-MCs display substantial release of β-hexosaminidase and CD63 upregulation after exposure to classic MC activators IgE/anti-IgE or C48/80. ( A ) Degranulation induced by various stimuli and ( B ) CD63 upregulation after successfully induced degranulation. PM-MCs generated from 3 different donors (orange, red, and blue bars) and control BC-MCs (green bars) were sensitized overnight with human-IgE and stimulated with anti-IgE, C48/80, or A23187, as explained in . Unstimulated MCs (NC) served as controls to correct for background degranulation. Each bar shows the mean ± standard deviation of 6 technical replicates per donor and stimulation condition. Gating for the data shown in ( B ) is explained in . The Figure was created with GraphPad Prism 9.2.0 software.

Article Snippet: Following IgE sensitization, cells were washed with Tyrode’s Salts buffer (Sigma-Aldrich) supplemented with 1.5 mM CaCl 2 and incubated for 45 min of incubation at 37 °C with the following stimuli: 50 μg/mL compound 48/80 (C48/80, Merck via Sigma-Aldrich) 1 μg/mL mouse anti-human IgE (clone G7-26, BD Biosciences, Drachten, The Netherlands) or 1 μM calcium ionophore A23187 (Merck via Sigma-Aldrich).

Techniques: Generated, Control, Standard Deviation, Software

Journal: medRxiv

Article Title: Altered neutrophil G-protein receptor signalling linked to impaired chemotaxis and increased ROS and NET production in older people with frailty

doi: 10.64898/2025.12.16.25342352

Figure Lengend Snippet: (A) ROS production in response to fMLP in neutrophils primed with GM-CSF (5ng/mL), TNFa (10ng/mL) or unprimed (UP). (B) NET production in response to PMA (0.1mg/mL), A23187 (3.8mM ), LPS (1 or 10 ng/mL) or unstimulated (cells only). Purple = FR (n=5-10), Pink = RA (n=5-9), Orange = HO (n=5-10), Green = HY (n=5-7). Analysis by ANOVA (*p<0.05, **p<0.01, ***p>0.001). (C) Representative microscopy images of NETs on coverslips (Blue = DNA, Red = MPO, Green = neutrophil elastase). Scale bar 50μM.

Article Snippet: Cells were allowed to settle for 15min prior to addition of LPS from P. aeruginosa (1 or 10ng/mL, Merck), PMA (0.1mg/mL, Merck), or A23187 (3.8μM, Merck) along with an untreated control.

Techniques: Microscopy