Proteome Profiler Human Phospho-RTK Array Kit (R&D Systems, #ARY001B) was used to detect the potential activation of RTK signals by hRNase 1 treatment. All procedures were performed according to the manufacturer’s instruction. Briefly, capture antibodies for specific RTKs were spotted in duplicate onto nitrocellulose membranes, prepared with the kit. Cell lysates (600 µg) were incubated with the array membrane at 4 °C overnight. After washing away unbound material, proteins in cell lysates containing phosphorylated tyrosine residues bound to the capture antibodies of RTKs were detected by a pan anti-phospho-tyrosine antibody conjugated to HRP. Last, the binding signal was measured by using chemiluminescent detection reagents and ImageQuant LAS 4010 (GE Healthcare).
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