Figure 3.

In vitro assays for RNase activity of PNGM-1 on total human cell RNA. Total RNA (0.3 μg) of the human cell line KMM-1 was incubated with or without 15 μM PNGM-1 in the presence of 10 mM MgCl₂ (A) or MnCl₂ (B), at 37°C for 60 min, and analyzed by microfluidics-based automated electrophoresis. Input RNA without PNGM-1 in the absence of any additional metal ions is shown in (C). An electropherogram and a gel image for each sample are shown. The peaks around 22 and 25 s denote the 25-nt marker (green line on the gel image) and approximately 100-nt RNAs including tRNA, 5S rRNA, and 5.8S rRNA, respectively. 18S, 18S rRNA (pink line); 28S, 28S rRNA (blue line); FU, arbitrary fluorescence units; RIN, RNA integrity number. Experiments in (A)-(C) were repeated at least three times and were reproducible.