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Bam HI

A. XbaI. 10xHis. FHA domain. BamHI. Phosphate domain. Kinase domain. Bam HI. 5’-. -3’. C. F10. B. Lysate. FL. W1. W2. F1. F2. F3. F4. F5. F6. F7. F8. F9. 62. Bam HI/XbaI. Bam HI. Marker. 47.5. 32. 10..0K. 25. 8.0K. 6.0K. 5.0K. 4.5K. 16.5. 4.0K. 3.5K. 3.0K.

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Bam HI

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  1. A XbaI 10xHis FHA domain BamHI Phosphate domain Kinase domain Bam HI 5’- -3’ C F10 B Lysate FL W1 W2 F1 F2 F3 F4 F5 F6 F7 F8 F9 62 Bam HI/XbaI Bam HI Marker 47.5 32 10..0K 25 8.0K 6.0K 5.0K 4.5K 16.5 4.0K 3.5K 3.0K 2.5K 2.0K D antiPNK 1.5K 1.0K monoAb PolyAb 0.5K 0.25K 16.5K Supplemental figure 1. Cloning and purification of PNKP FHA domain. (A) Diagram of the PNKP cDNA; (B) serial restriction enzyme digestion of PNKP cDNA with BamHI and XbaI; (C) chromatography of His-tagged FHA domain on Ni-NTA column; (D) western blot analysis showing both monoclonal and polyclonal antibodies of PNKP can detect FHA domain.

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