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. 2015 Oct 12;6(37):40141–40157. doi: 10.18632/oncotarget.5653

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Copyright: © 2015 Mazzacurati et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

PMC Copyright notice

A. PIM1L and PIM1S were expressed from a viral promoter in BaF3-JAK2-V617F cells. Cell lysates were immunoblotted for PIM1, actin as a loading control, and MDM2 for a proteasome inhibitor control. The proteasome inhibitor bortezomib was utilized to stabilize PIM1 expression for easier detection of PIM1S. The exogenous PIM1 proteins were FLAG-tagged thus increasing their molecular weight and slowing their mobility in SDS-PAGE compared to endogenous PIM1 proteins. Mobility of endogenous and exogenous PIM1 proteins are indicated with arrows. B. These cells from A., along with vector control, were cultured with DMSO or ruxolitinib (0.5 μM and 1.0 μM) and total viable cells were determined over time by trypan blue exclusion. Similar results were obtained in three independent experiments.