(A) Cell Cycle Analysis by FACS. Summary of previously published data (14).
(B) Expression of cyclin D1 and Cyclin-Dependent Kinase inhibitors in B3CD treated NB cells. Expression of cyclin D1, p21 and p27 cell cycle regulators in B3CD and vehicle treated SMS-KCNR or SH-SY5Y cells were analyzed by western blotting of lysates with the appropriate primary and secondary antibodies in combination with a chemiluminescence detection system as described in (Materials and Methods). As an internal standard for equal loading (50 μg total cell protein/lane) blots were probed with an anti-GAPDH antibody.
(C) Semi-quantitative RT-PCR of cyclin D1 and Cyclin-Dependent Kinase inhibitors in B3CD treated NB cells. Total RNA was isolated from SMSK-CNR or SH-SY5Y cells treated with 0, 1 and 3 μM of B3CD for 48 h. RT PCR was carried out as described (Material and Methods).