Fig. 1.

Ang II–evoked TRPC5 and TRPC6 activation contributes to intracellular Ca²⁺ transients in podocytes. Fluo-3 Ca²⁺ imaging in AT1R podocytes revealed Ca²⁺ transients in response to Ang II (500 nM) with (2 mM) or without Ca²⁺ (0 mM + 5 mM EGTA). Ionomycin(Ion)was added to the bath at the end of the experiment. (A to C) Normalized fluorescence amplitude in response to Ang II application (F) in podocytes expressing either scrambled (Scr) (A), C5 (B), or C6 shRNA (C). (D) Group data summary (mean ± SD). Gene silencing of TRPC5 (n = 34) and TRPC6 (n = 29), but not of TRPC1 (n = 15), TRPC4 (n = 26), or TRPC7 (n = 42), significantly decreased the amplitude of Ca²⁺ transient. (E to G) Representative traces in Ca²⁺ add-back experiments. A proportion of AT1R podocytes responded to a second application of Ang II within the 10-min recording period. (F) Gene silencing of TRPC5 (n = 5) or TRPC6 (n = 18) compared to Scr shRNA control (n = 16), significantly reduced Ca²⁺ influx in response to Ang II. (H) Group data in Scr controls, C5, and C6 knockdown cells (mean ± SE) (^##P < 0.01).