Figure 2.

Primary cell culture from T. castaneum brains. (A) Cultured brain ells 3 h after dissociation. (B) Cultured brain neurons labeled with anti-horseradish peroxidase (anti-HRP) (green) and DAPI (white) after 6 days in vitro. Inset shows regenerated neurites with optimized brightness and contrast. (C) Cultured brain cells after 1 day in vitro. Left and middle: DAPI labeling. Right and middle: anti-HRP immunofluorescence. *Intact neurons, ^#dying neuron, ^onucleus of dead or dying cell, ⁺intact glial cell. (D) DAPI labels heterochromatin DNA structures in nuclei of intact cells (*) and condensed nuclei with largely fragmented DNA of dead or dying cells (o). (E) Anti-cleaved caspase-3 immunofluorescence (green) in the cytoplasm of apoptotic cells. Dead cells (condensed DAPI-labeled nuclei) lose their cleaved caspase-3 associated fluorescence after disintegration of cellular membranes. Activated caspase-3 is absent from the cytoplasm of intact cells with discontinuously DAPI-labeled nuclei.