American Society of Hematology

Figure 2

$Figure 2. Cytoplasmic p27 promotes survival of CML cells. (A) p27 immunoblotting was performed on the nuclear and cytoplasmic fractions of exponentially growing CML cell lines. K562 and KYO-1 cells show predominantly cytoplasmic p27, whereas Mo7ep210 cells show predominantly nuclear p27. (B) K562, KYO-1, and Mo7ep210 cells were infected with p27 or scrambled shRNA lentivirus and total p27 expression was determined by immunoblot analysis. (C) Cells infected with p27 or scrambled shRNA were analyzed for apoptosis using Annexin V staining. *P < .050; **P < .010. (D) Effect of cytoplasmic-only p27 (p27ΔNLS) on cell growth of CML cells. p27−/− MEFs were transduced with Flag-p27ΔNLS and localization was validated by IF analysis using anti-Flag antibody (M2; Sigma-Aldrich). DAPI was used for nuclear staining. K562 and Mo7ep210 cells were transduced with p27ΔNLS or empty vector control and the effect on cell growth was measured over time.$

Cytoplasmic p27 promotes survival of CML cells. (A) p27 immunoblotting was performed on the nuclear and cytoplasmic fractions of exponentially growing CML cell lines. K562 and KYO-1 cells show predominantly cytoplasmic p27, whereas Mo7ep210 cells show predominantly nuclear p27. (B) K562, KYO-1, and Mo7ep210 cells were infected with p27 or scrambled shRNA lentivirus and total p27 expression was determined by immunoblot analysis. (C) Cells infected with p27 or scrambled shRNA were analyzed for apoptosis using Annexin V staining. *P < .050; **P < .010. (D) Effect of cytoplasmic-only p27 (p27^ΔNLS) on cell growth of CML cells. p27^−/− MEFs were transduced with Flag-p27^ΔNLS and localization was validated by IF analysis using anti-Flag antibody (M2; Sigma-Aldrich). DAPI was used for nuclear staining. K562 and Mo7ep210 cells were transduced with p27^ΔNLS or empty vector control and the effect on cell growth was measured over time.

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