Fig 6. Delta-cell composition of islets after 7 days of tissue culture in control media (—) or media supplemented with 4 different doses of necrostatin-1 (25, 50, 100, or 200 μM of Nec-1) on day 3.

3000 islet equivalents were dissociated into single cells using Accutase, stained with 7-amino-actinomycin D (7-AAD) viability dye and fluorescein-isothiocyanate-conjugated anti-somatostatin antibody, and analyzed by flow cytometry to quantify the proportion of delta cells in islets. Somatostatin-positive delta cells were gated on 7-AAD-negative live cells. A) Representative flow cytometry plots of somatostatin staining. B) Percentage of somatostatin-positive delta cells. n = 6 for each group. *p < .05. **p < .01. Data are expressed as mean ± SEM.