Figure 7.

The enhanced survival capacity of S1P₁-expressing cells requires new protein synthesis and is lost upon Mcl-1 downregulation. (a) Control and S1P₁-expressing CCL39 cells were switched to serum-free medium (SF) for 16 h before washing and incubation in SF for the indicated times in the presence of protein synthesis inhibitor emetine (100 μM). Cell extracts were then prepared for fractionation via SDS-PAGE and subsequent immunoblotting with the indicated antibodies. Quantitation of cleaved caspase-3 and Bim expression normalised to GAPDH in control and S1P₁-expressing CCL39 cells is presented as mean values±S.E. for n=3 separate experiments. *P<0.05 versus CCL39 control cells. (b) Control and S1P₁-expressing CCL39 cells were switched to SF medium for 16 h before washing and incubation in SF for the indicated times in the presence of protein synthesis inhibitor emetine as described in (a). Cell extracts were then prepared for fractionation via SDS-PAGE and subsequent immunoblotting with the indicated antibodies. Quantitation of Mcl-1expression normalised to GAPDH in control and S1P₁-expressing CCL39 cells is presented as mean values±S.E. for n=3 separate experiments. *P<0.05 versus CCL39 control cells