Supplemental Table S 1
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چکیده
Plasmids and cloning cDNA for human USP4 and USP8 was a gift from Hidde Ploegh and cDNA for USP39 was a gift from R. Medema. USP4CD (aa 296-954) and the D1 fragment (aa 296-490) of human USP4, USP39CD (aa 219-565) and USP8CD (aa 771-1118) were cloned using ligation independent cloning into pET-46 Ek/LIC vector (Novagen). The D2 fragment (aa 766-932) of USP4 was cloned into the pET-NKI b/3C (Luna-Vargas, in preparation). The fused USP4-D1D2 was created by inserting aa 353-359 of USP7 (SIKGKNN) between residues Leu479 and Leu777. The USP4 insert (aa 483-765), Ubl domain (aa 483-571) and Ub were cloned into pGEX-6P-1 (GE Healthcare). The USP8CD mutant was generated by site-directed mutagenesis of the catalytic cysteine (C786A).
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