Bi et al. 10.1073/pnas.0703805104. |
Fig. 6. The effect of colchicine on neuronal viability. (A) The morphology of dorsal root ganglion (DRG) neurons and axons in the central and side compartments before (Upper) and after (Lower) colchicine treatment. (B) The expression of GAP43 in axons of DRG neurons monitored by immunohistochemistry. BF, bright field; -Colch, without colchines treatment; +Colch, with colchicine treatment. (C) Statistical analysis of the number of trypan blue-stained DRG neurons with (+) or without (-) colchicine treatment (n = 4, P > 0.025)
SI Text
Viability assay.
Rat dorsal root ganglion neurons were seeded in 6-cm culture plates [coated with poly(D-lysine)] for 5 days and treated with, or without, 10 mg/ml colchicine for 1 h. Neurons were detached with 0.05% trypsin-EDTA treatment for 5 min at 37°C after 1X PBS washing. Trypsin-EDTA was removed and cells were suspended in a dorsal root ganglion neuron culture medium containing 10% FBS. The cell suspension was diluted, 1:1, with 0.4% trypan blue. The number of stained cells was scored using a hemocytometer. The results were analyzed by the paired Student's t test.