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. 2014 Jan;28(1):395–407. doi: 10.1096/fj.13-235382

Figure 4.

Figure 4.

Akt3 controls CRM-1 protein stability. A) Real-time PCR of mRNA isolated from HUVECs transfected with a scrambled control (SCR) or Akt3 RNAi (Akt3i) assessed for CRM-1 expression and shown as relative to S26. B) Western blot analysis of CRM-1 expression in total lysates derived from HUVECs transfected with SCR or Akt3i and treated cycloheximide for the times indicated. α-Tubulin shown as loading control. C) Quantification of relative CRM-1 expression following 24 h of cycloheximide treatment in HUVECs transfected with SCR or Akt3i. *P < 0.05. D) Diagram of CRM-1 indicating RanGTP and cargo-binding domains and the binding site of leptomycin B. Sequences showing the wild-type and mutated serine 1054 residues are shown. E) Western blot analysis of FLAG expression in HUVECs transfected with either wild-type or S1054A mutant FLAG-tagged CRM-1 constructs shown in C and treated with cyclohexamide for the times indicated. F) Quantification of relative CRM-1-FLAG expression following 24 h of cycloheximide treatment in HUVECs transfected with either wild-type or S1054A mutant FLAG-tagged CRM-1 plasmid. *P < 0.05.