Mutations in TRPC6 lead to increased intracellular calcium ([Ca²⁺]_i) signalling and a phenotype of familial nephrotic syndrome, affecting the glomerular podocyte (1,2). In addition, in some acquired glomerulopathies increased podocyte TRPC6 expression is associated with the re-localisation, or downregulation of nephrin (3,4,5,6), a cell adhesion molecule and podocyte marker associated with the slit diaphragm. We sought to determine how podocytes are uniquely affected by TRPC6 mis-function and the role of nephrin using human conditionally immortalised podocytes (ciPods) and nephrin-deficient (ND) ciPods, previously characterised (7,8). Cells were loaded with Fura2-AM, Methyl β cyclodextrin (MβCD), when used was added in the final 30mins of loading. Cells, incubated in Krebs’ solution containing 1.5mM or 200nM Ca²⁺, were stimulated with 200μM flufenamic acid (FFA) in the presence or absence of thapsigargin (TG). [Ca²⁺]_i was proportional to the fluorescence intensity ratio (R_norm). ciPods transfected with Ad-virus containing wild type and dominant negative TRPC6 constructs verified the specificity of FFA to TRPC6. 200μM FFA increased the R_norm in ciPods in 1.5mM [Ca²⁺]_o (1.44±0.11 fold increase, p≤0.01 paired t-test) and surprisingly in 200nM [Ca²⁺]_o (1.58±0.01 fold increase, P≤0.01, paired t-test), but not after store-depletion (R_norm1; 1.22±0.06 fold increase) in contrast to an increase in ND ciPods after store-depletion (1.6±0.06 fold increase, p≤0.05, paired t-test). Pre-incubation of ciPods with 1mM MβCD inhibited the response to FFA in 1.5mM [Ca²⁺]_o as expected (R_norm 1.07±0.03 fold increase), yet also blocked the response in 200nM [Ca²⁺]_o (R_norm1.19±0.01 fold increase), verifying TRPC6 function as a store calcium release channel. In conclusion, FFA specifically activates TRPC6 in ciPods. In these cells TRPC6 can uniquely act as a store calcium-release channel, although only in the presence of nephrin. The activation of TRPC6 is lipid raft dependent, whether in the plasma membrane or on internal stores.