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. 2002 Feb 18;156(4):703–713. doi: 10.1083/jcb.200110063

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Copyright © 2002, The Rockefeller University Press

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Figure 8. — Photobleaching reveals reduced monomer exchange in KelY627A and src64 ^Δ17 ring canals. FRAP experiments with GFP–actin were used to measure actin dynamics in kelch mutant ring canals, containing either wild-type (P[ORF1]) or mutant (P[kelY627A]) rescuing transgenes, and in src64 ^Δ ¹⁷. (A) A representative time series of FRAP in a ring canal expressing wild-type Kelch (P[ORF1]). The box outlined at 25 s represents the area photobleached by the laser. By 168 s, all of the fluorescence had returned. (B) A similar time series of FRAP in a ring canal expressing nonphosphorylatable Kelch (P[kelY627A]). The area bleached at 25 s took 420 s to recover. (C) A FRAP assay of src64^Δ17, where the area bleached at 25 s took 435 s to recover. Bars, 5 μm. (D) A graph representing FRAP of 21 P[ORF1] (▪), 23 P[kelY627A] (▴), and 15 src64 ^Δ ¹⁷ (×) stage 10A ring canals with error bars representing the standard deviation. P[kelY627A] and src64 ^Δ ¹⁷ ring canals take more than three times as long to recover to prebleach levels as P[ORF1] ring canals.