Figure 5
Figure 5. The ATIC activity is enhanced by NPM-ALK in HEK-293T-Rex cells. (A) NPM-ALK inducible HEK-293T-Rex cells were induced with tetracycline with and without CEP14083 ALK inhibitor. p-NPM-ALK was determined by Western blotting using anti–p-ALK antibodies. ATIC and LDH enzymatic activities were evaluated; *P < .001. (B) HEK-293T-Rex cells containing an inducible ATIC-ALK construct were induced with tetracycline and/or with CEP14083. Western blotting analysis confirmed ALK expression and phosphorylation. Enzymatic activity of ATIC and LDH was evaluated. (C) Cell lysates of transiently transfected HEK-293T cells (NPM-ALK, NPM-ALKK210R, ATIC-ALK, TPR-Met, c-myc, and the self-activating form of Ras, Ras V12) were tested for their ATIC and LDH enzymatic activity. *P < .001, **P < .05. (D) Cell lysates from transiently transfected HEK-293T cells with NPM-ALK, NPM-ALKK210R, and/or a Ras DN, respectively, were tested for their ATIC and LDH enzymatic activity.

The ATIC activity is enhanced by NPM-ALK in HEK-293T-Rex cells. (A) NPM-ALK inducible HEK-293T-Rex cells were induced with tetracycline with and without CEP14083 ALK inhibitor. p-NPM-ALK was determined by Western blotting using anti–p-ALK antibodies. ATIC and LDH enzymatic activities were evaluated; *P < .001. (B) HEK-293T-Rex cells containing an inducible ATIC-ALK construct were induced with tetracycline and/or with CEP14083. Western blotting analysis confirmed ALK expression and phosphorylation. Enzymatic activity of ATIC and LDH was evaluated. (C) Cell lysates of transiently transfected HEK-293T cells (NPM-ALK, NPM-ALKK210R, ATIC-ALK, TPR-Met, c-myc, and the self-activating form of Ras, Ras V12) were tested for their ATIC and LDH enzymatic activity. *P < .001, **P < .05. (D) Cell lysates from transiently transfected HEK-293T cells with NPM-ALK, NPM-ALKK210R, and/or a Ras DN, respectively, were tested for their ATIC and LDH enzymatic activity.

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