FIG. 1.
Establishment of primary mouse thyrocyte cultures and assessment of NaI-induced apoptosis. NOD.H2h4 thyrocytes were expanded in culture for 10 days, and their purity was assessed by a two-step immunofluorescence assay using FITC-labeled goat anti-mouse IgG, as described in the Materials and Methods. (A) Thyrocytes incubated with NOD.H2h4 serum containing high titers of Tg-specific IgG. (B) Thyrocytes incubated with control serum from healthy CBA/J mice. Appearance of NOD.H2h4 thyrocytes following exposure to apoptotic/necrotic stimuli. (C) Healthy thyrocytes (blue staining of nucleus with Hoechst 33342). (D) Early apoptotic thyrocyte cultured in 10−5 M NaI (green fluorescent staining of cytoplasm with FAM-FLICA poly caspase reagent). (E) Late apoptotic thyrocyte cultured in 10−5 M NaI (cytoplasmic stain with FAM-FLICA and propidium iodide). (F) Necrotic thyrocyte following exposure to 90% ethanol for 30 sec (cytoplasmic stain with propidium iodide).