Sustained mechanical stresses (e.g. high blood pressure) damage the renal glomerulus resulting in proteinuria. Podocytes express the canonical transient receptor potential 6 (TRPC6) channel, the activity of which is modulated by receptor and mechanical stimulations in a complex manner. However, the implication of this modulation for the glomerular barrier function is rather poorly elucidated. To address this point, we carried out the albumin-permeation assay using immortalized mouse podocytes stably expressing the wild-type (wt) TRPC6 or its gain-of-function mutant associated with focal segmental glomerulosclerosis (FSGS), M131T. Differentiated podocytes were grown on cell-culture insert membranes (pore size: 0.4micrometer). Compared to unstimulated or angiotensin II (Ang II) alone, simultaneous stimulation with Ang II and a membrane-expanding agent 2,4,6-trinitrophenol (TNP) reduced the leak of FITC-labelled albumin across the membranes in both wt-TRPC6- and M131T-expressing podocytes. SAR7334, a potent TRPC6-specific inhibitor, increased the albumin leak, the effect being more prominent in the latter. These results suggest that TRPC6 activation by simultaneous receptor and mechanical stimulations may reinforce the filtration barrier function mediated by podocytes, and this may be impaired by the FSGS-associated mutation M131T.