Figure 6. Panobinostat induced p21^Waf1/Cip1 expression.

FaDu cells were synchronised by double thymidine block and released in medium containing 100 nM panobinostat or excipient control. Cells were harvested at the stated time points post release and RNA (A) and protein (B and C) harvested as described in the materials and methods. (A) p21^Waf1/Cip1 mRNA expression was assessed by qRT-PCR. The data shown represent p21^Waf1/Cip1 mRNA levels normalised to 0 hour time point and are shown as the mean and standard deviation of three independent experiments. (B) 60 μg protein from each extract was separated by SDS-PAGE on two separate gels runs using identical conditions and p21^Waf1/Cip1 (upper panel) and β-actin (lower panel) protein levels were assessed by Western blot. Full-length digital images of these western blots can be viewed in the supplementary information (supplementary figure 5). (C) p21^Waf1/Cip1 protein levels from three independent experiments were quantified using ImageGauge v4.21 software and normalised to β-actin levels from the same experiment. The data shown represent the mean and standard deviation.