We carried out a yeast two-hybrid screen to identify proteins that interact with large conductance (BK_Ca-type) Ca²⁺-activated K⁺ channels encoded by the Slo1 gene. Nephrin, an essential adhesion and scaffolding molecule expressed in podocytes, emerged in this screen. The Slo1-nephrin interaction was confirmed by co-immunoprecipitation from brain and kidney, and from HEK293T cells expressing both proteins, and by GST pull-down assays. We detected nephrin binding to the Slo1_VEDEC splice variant, which is typically retained in intracellular stores, and to the 4 subunit. However, we did not detect significant binding of nephrin to the Slo1_QEERL or Slo1_EMVYR splice variants. Co-expression of nephrin with Slo1_VEDEC increased expression of functional BK_Ca channels on the surface of HEK293T cells, but did not affect steady-state surface expression of the other C-terminal Slo1 variants. Nephrin did not affect the kinetics or voltage-dependence of channel activation in HEK293T cells expressing Slo1. Stimulation of Slo1_VEDEC surface expression in HEK293T cells was also observed by co-expressing a small construct encoding only the distal C-terminal domains of nephrin that interact with Slo1. Reduction of endogenous nephrin expression by application of siRNA to differentiated cells of an immortalized podocyte cell line markedly reduced the steady-state surface expression of Slo1 as assessed by electrophysiology and cell-surface biotinylation assays. Nephrin therefore plays a role in organizing the surface expression of ion channel proteins in podocytes, and may play a role in outside-in signaling to allow podocytes to adapt to mechanical or neurohumoral stimuli originating in neighboring cells.