Fig. 1.

Identification of Myo1c as a Neph1 binding protein. (A) Eluates from Neph1 affinity and control columns were separated on SDS-PAGE gels and subjected to mass spectrometry. Protein bands containing ZO-1, Neph1, and Myo1c are highlighted. (B) Normalized spectrum values derived by using Scaffold software (Proteome Software, Inc., Portland, OR). (C) The same eluate was probed with Neph1 and Myo1c antibodies to confirm the results from the mass spectrometry analysis. Various myosin I proteins are expressed in human podocytes. (D) RT-PCR of differentiated and undifferentiated cultured human podocytes with the indicated primers detects various myosin family members. Mr, marker. (E) Cultured human podocytes and rat glomerular lysates were Western blotted (WB) with various class I myosin antibodies.