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. 2016 Jun 9;7(29):45249–45262. doi: 10.18632/oncotarget.9932

Figure 3. Pyrvinium pamoate activates the AMPK/importin α1 cascade.

Figure 3

(A) Pyrvinium pamoate activates AMPK and decreases HuR cytoplasmic abundance in a dose-dependent manner. (B) Immunofluorescence assays shows that pyrvinium pamoate inhibits doxorubicin-triggered cytoplasmic translocation of HuR by activating the AMPK pathway. 5637 cells were treated with doxorubicin (360 nmol/L) for 12 h, followed by indicated treatments (100 nmol/L pyrvinium pamoate, 10 μmol/L compound C and 2 mmol/L AICAR) for an additional 6 h. Immunofluorescence staining for HuR was performed (magnification, 40×). (C) Pyrvinium pamoate improves the interaction of HuR and importin α1. Cells transfected with equivalent amount of flag-pcDNA3.1 and myc-importin α1 served as the negative control. (D) Importin α1 is required for pyrvinium pamoate-mediated HuR nuclear import. 5637 cells were transfected with pcDNA3.1 or importin α1 (wild-type) or importin α1 (K22R/S105A, dual-site mutation). CE, cytoplasmic extracts; WCE, whole-cell extracts; Dox, doxorubicin; PP, pyrvinium pamoate; CC, compound C; AICAR, sodium azide, 5-amino-imidazole-4-carboxamide riboside.