Figure 2
SOCS3 enhances JAK2 V617F phosphorylation and fails to be degraded. (A) 293T cells were transfected with combinations of EPOR, JAK2, or JAK2 V617F, and FLAG-tagged SOCS1, SOCS2, or SOCS3 as indicated. Cells were treated for 30 minutes with EPO (50 U/mL) (+EPO, top 3 panels) or left untreated (−EPO, bottom 3 panels). Lysates were immunoblotted with phosphotyrosine antibody then stripped and reprobed with JAK2 antibody (top panels) or anti-FLAG to detect SOCS proteins. (B) Cell lysates from panel A were simultaneously analyzed for SOCS protein and mRNA levels. Whole-cell lysates were immunoblotted with FLAG antibody to detect SOCS proteins, while RT-PCR was performed on the total RNA using primer pairs specific for the SOCS genes, and products were separated by 2% agarose gel electrophoresis and visualized by ethidium bromide (bottom panels).

SOCS3 enhances JAK2 V617F phosphorylation and fails to be degraded. (A) 293T cells were transfected with combinations of EPOR, JAK2, or JAK2 V617F, and FLAG-tagged SOCS1, SOCS2, or SOCS3 as indicated. Cells were treated for 30 minutes with EPO (50 U/mL) (+EPO, top 3 panels) or left untreated (−EPO, bottom 3 panels). Lysates were immunoblotted with phosphotyrosine antibody then stripped and reprobed with JAK2 antibody (top panels) or anti-FLAG to detect SOCS proteins. (B) Cell lysates from panel A were simultaneously analyzed for SOCS protein and mRNA levels. Whole-cell lysates were immunoblotted with FLAG antibody to detect SOCS proteins, while RT-PCR was performed on the total RNA using primer pairs specific for the SOCS genes, and products were separated by 2% agarose gel electrophoresis and visualized by ethidium bromide (bottom panels).

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal