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Figure 8


Fig. 8. Knockdown of eIF3k promotes keratin-dependent sequestration of active caspase 3 in the insoluble compartment of cells. SW13-K8/K18 (A) or SW13 (B) cells stably expressing eIF3k siRNA or control siRNA as described in Fig. 7E and Fig. 7A, respectively, were irradiated with or without UV at 0.015 J/cm2 and then cultivated for 7 hours. Cells were extracted with lysis buffer containing 0.5% Triton X-100, and soluble and insoluble fractions were separately collected as described in the Materials and Methods. Equal volumes of the soluble and insoluble fractions were mixed as the total lysates (Total). One fifteenth of each fraction and total lysates were resolved by SDS-PAGE and then analyzed by western blot with antibodies as indicated. CrkII and K8 (or Lamin A) were used as the reference protein for soluble and insoluble fractions, respectively.





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