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. 2020 Mar 20;25(6):1414. doi: 10.3390/molecules25061414

Figure 4.

Figure 4

Comparison of the DNase activity of full-length hDicer (hDcr) and the PAZ domain deletion variants ΔPAZ_hDcr and ΔPPC_hDcr on short ssDNA substrates. (A,B; left panels) The results of the DNA-cleavage assays involving: 21-nt ssDNA (A; left panel) and 32-nt ssDNA (B; left panel), and increasing amounts of the respective protein (hDcr, ΔPAZ_hDcr or ΔPPC_hDcr): 12.5, 25, 50, 75 nM (represented by a triangle). (A,B; right panels) The results of the time-dependent cleavage of 21-nt ssDNA (A; right panel) and 32-nt ssDNA (B; right panel). Triangles represent time points: 15 min, 30 min, 1h. Reactions were carried out with 50 nM protein (hDcr, ΔPAZ_hDcr or ΔPPC_hDcr). (C-) controls incubated without a protein. (C+EDTA) controls incubated without a protein but with 25 mM EDTA. (+EDTA) supplementation of the reaction buffer with 25 mM EDTA. (T1) G-ladder generated with RNase T1.