960378.fig.003a
(a)
960378.fig.003b
(b)
Figure 3: Modulation of IGF-induced Akt phosphorylation in response to 1, 25-D3 treatment in MCF-7 and MCF-7/VDR cells. (a) MCF-7 and MCF-7/VDR cells were treated with 100 nM 1, 25-D3 or 30 nM IGF-I, alone or in combination in serum-free medium. Cells were also cultured in medium supplemented with 2% serum as a control. After 6 days of treatment, cell viability was estimated by neutral red assay. * and ** are statistically significant compared to control. Means of 3 separated experiments are shown ( ). (b) (i) MCF-7 and MCF-7/VDR cells were treated with 100 nM 1, 25-D3 or 30 nM IGF-I, alone or in combination, in serum-free medium. After 5 days of treatment, whole cell extracts were prepared and analysed on a phospho-MAPK antibody array (R&D Systems, UK) following manufacturer’s instruction. (ii) Densitometric analysis of Akt phosphorylation. Data shown are means of 3 replicates, significantly different from parental cells. ** .