Fig. 7.
Fig. 7. Expression of TEL-JAK2 results in constitutive activation of SAPK/JNK activity. / Ba/F3 and Ba/F3-TEL-JAK2 cell lines were depleted of cytokine and then stimulated in the presence (+) or absence (−) of IL-3. SAPK was immunoprecipitated, and an in vitro kinase assay was performed using GST–c-Jun as substrate. Reaction mixtures were resolved by SDS-PAGE, and radiolabeled proteins were detected by PhosphorImager analysis (upper panel). Total SAPK was detected by IB with a SAPK antibody (lower panel). (B) A total of 2000 Ba/F3 cells (closed squares) and Ba/F3-TEL-JAK2(5-19) cells in the absence of IL-3 (open circle) or Ba/F3-TEL-JAK2(5-19) in the presence of IL-3 (closed circles) were seeded into 96-well plates containing media of increasing SP600125 concentration. The cells were incubated for 48 hours at 37°C prior to the addition of XTT and phenazine methosulfate (PMS). Absorbance was read at 450 nm following 4 hours of incubation at 37°C.

Expression of TEL-JAK2 results in constitutive activation of SAPK/JNK activity.

Ba/F3 and Ba/F3-TEL-JAK2 cell lines were depleted of cytokine and then stimulated in the presence (+) or absence (−) of IL-3. SAPK was immunoprecipitated, and an in vitro kinase assay was performed using GST–c-Jun as substrate. Reaction mixtures were resolved by SDS-PAGE, and radiolabeled proteins were detected by PhosphorImager analysis (upper panel). Total SAPK was detected by IB with a SAPK antibody (lower panel). (B) A total of 2000 Ba/F3 cells (closed squares) and Ba/F3-TEL-JAK2(5-19) cells in the absence of IL-3 (open circle) or Ba/F3-TEL-JAK2(5-19) in the presence of IL-3 (closed circles) were seeded into 96-well plates containing media of increasing SP600125 concentration. The cells were incubated for 48 hours at 37°C prior to the addition of XTT and phenazine methosulfate (PMS). Absorbance was read at 450 nm following 4 hours of incubation at 37°C.

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal