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Fig. 7. TRPC1 activity depends on caveolin-3 expression and Src phosphorylation. C2 myoblasts were transfected with TRPC1-CFP alone or in combination with caveolin-3–YFP. Cells were loaded with Fura Red and imaged using scanning confocal microscopy. During imaging, cells were incubated with H₂O₂ and changes in Ca²⁺ levels were measured. (A) Fura Red is excited using the 458 nm and 488 nm laser lines (upper graph). Relative Ca²⁺ concentration is represented by the ratio of Fura Red emission upon 458 nm and 488 nm excitations (ratio=Em_458nm/Em_488nm; lower graph). (B) C2 myoblasts were co-transfected with TRPC1-CFP and caveolin-3–YFP, loaded with Fura Red, and imaged. Increase in Ca²⁺ levels, compared with control (non-transfected cells; thick black line), was detected in such cells upon H₂O₂ incubation and this increase was prevented by PP2 incubation prior to H₂O₂ treatment (dotted line). In cells expressing TRPC1-CFP only (no caveolin-3–YFP), no Ca²⁺ increase was induced by H₂O₂ incubation (thin grey line). (C) Pooled data showing the increase in Ca²⁺ influx in TRPC1–caveolin-3-expressing cells upon Src activation (by H₂O₂). Graph represents mean ± s.e.m. of a minimum of 20 cells from four independent experiments. ^***P<0.001.

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