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Double-labeling immunofluorescence for pSmad2/3 and α-SMA in cultured pulp tissue without protease inhibitors. Nuclei (DAPI, blue), pSmad2/3 (green) and α-SMA (red) are shown in uncultured normal pulp at 2, 5 and 7 days of culture. Nuclear accumulation of pSmad2/3 is detectable at 2 days (arrows); thereafter, pSmad2/3 and α-SMA double-positive cells are evident at 5 days (arrows). α-SMA single-positive cells are abundantly observed at 7 days (arrows). Scale, 10 µm.
