American Society of Hematology

Figure 1.

$Figure 1. Expression of NPM-ALK in inducible and constitutive Ba/F3 expression systems. (A) Expression of the fusion protein NPM-ALK (80 kDa) in TonBaF.1-NPMALK cells 24 hours and 48 hours after the addition of 2 μg/mL doxycycline. Lanes A, B, and C correspond to 3 independent retroviral transductions of TonBaF.1 cells with pRev-TRE-Hyg-NPM-ALK. Uninduced samples were harvested from cells starved of IL-3 for 17 hours. +Dox 24-hour and +Dox 48-hour samples were harvested from cells starved of IL-3 for 17 hours and induced with 2 μg/mL doxycycline for 24 hours and 48 hours, respectively. Whole-cell lysates were fractionated by SDS-PAGE and immunoblotted with NPM-ALK specific antibody. Cells derived from transduction C were used for subsequent analyses.(B) Expression of the fusion protein NPM-ALK in Ba/F3 cells stably transformed with NPM-ALK. Whole-cell lysates of MSCV and MSCV-NPM-ALK-transduced Ba/F3 cells were harvested, fractionated by SDS-PAGE, and immunoblotted with NPM-ALK-specific antibody.$

Expression of NPM-ALK in inducible and constitutive Ba/F3 expression systems. (A) Expression of the fusion protein NPM-ALK (80 kDa) in TonBaF.1-NPMALK cells 24 hours and 48 hours after the addition of 2 μg/mL doxycycline. Lanes A, B, and C correspond to 3 independent retroviral transductions of TonBaF.1 cells with pRev-TRE-Hyg-NPM-ALK. Uninduced samples were harvested from cells starved of IL-3 for 17 hours. +Dox 24-hour and +Dox 48-hour samples were harvested from cells starved of IL-3 for 17 hours and induced with 2 μg/mL doxycycline for 24 hours and 48 hours, respectively. Whole-cell lysates were fractionated by SDS-PAGE and immunoblotted with NPM-ALK specific antibody. Cells derived from transduction C were used for subsequent analyses.(B) Expression of the fusion protein NPM-ALK in Ba/F3 cells stably transformed with NPM-ALK. Whole-cell lysates of MSCV and MSCV-NPM-ALK-transduced Ba/F3 cells were harvested, fractionated by SDS-PAGE, and immunoblotted with NPM-ALK-specific antibody.

This Feature Is Available To Subscribers Only