Reduction of ROCK1 by MS-275 is dependant on protein expression. MTLn3 breast cancer cells were allowed to invade into HD matrices for 24 h before treatment with 1 μM and 3 μM MS-275, 10 μg/ml of cycloheximide (CHX) and both CHX and MS-275 for a further 24 h or cells were grown on LD and HD matrix for 72 h to total RNA and protein extraction. A, Real-time PCR results showed that MS-275 alone decreased ROCK1 expression but this was reversed in the presence of CHX. B, HD significantly suppressed Notch-1 expression at both mRNA and protein level compared to LD matrix and there was no change in p53 expression. C, MS-275 upregulated Notch1 levels and this was unaffected by CHX. Bars indicate standard error from two biological samples and experiments were performed two times. 1-way ANOVA followed by post-hoc Tukey’s test indicating significant difference at either * p < 0.05 or ** p < 0.01.
Raviraj et al. BMC Cell Biology 2012 13:12 doi:10.1186/1471-2121-13-12