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. 2002 Sep;161(3):875–883. doi: 10.1016/S0002-9440(10)64248-4

Figure 2.

Figure 2.

ALK is activated by the addition of coumermycin in vitro. Top: Myc-GyrB-ALK was transcribed and translated in vitro using reticulocyte lysates. Newly synthesized protein was then either mock-treated or treated with increasing concentrations of coumermycin for 30 minutes at 30°C. After treatment, the level of tyrosine phosphorylation of ALK was determined by immunoblotting with RC-20 anti-phosphotyrosine (α pTyr) antibody. Changes in the level of ALK tyrosine phosphorylation were seen suggesting coumermycin-induced autophosphorylation of GyrB-ALK. Bottom: A 5-μl aliquot of each transcription-translation reaction was analyzed by immunoblotting with 9E10 anti-Myc mAb (α Myc) as a control for the amount of Myc-GyrB-ALK present in each reaction.