Additional file 12:

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NPCD cells treated with 1 M NaOH solution. An increased in vesicle formation occurs, and later the vesicles fuse to the central vacuole immediately before tonoplast collapse. Prior to cell death there is a dramatic change in colour due to the effect of NaOH on anthocyanin in the underlying mesophyll. Also, nuclei disappeared and large amounts of precipitates are present in the vacuole immediately before tonoplast collapse. Note the swollen appearance of the chloroplasts. No PM retraction occurs. Approximately 2.5× playback speed.