Figure 4: β-Actin-GFP has a half-life of several hours at IHC stereocilia tips. | Nature Communications

Figure 4: β-Actin-GFP has a half-life of several hours at IHC stereocilia tips.

From: Length regulation of mechanosensitive stereocilia depends on very slow actin dynamics and filament-severing proteins

Figure 4

Explants of organ of Corti from P6 mice constitutively expressing β-actin-GFP in hair cells were cultured for 48 h and live cells were imaged by confocal microscopy. (a) Rectangular regions of IHC stereocilia bundles were photobleached and recovery of β-actin-GFP was monitored in the same bleached cells for times up to 24 h post-bleaching. The tips of stereocilia recovered β-actin-GFP fluorescence over time. (b) Cytochalasin D, which prevents actin polymerization at filament barbed ends, slowed the recovery of β-actin-GFP fluorescence at stereocilia tips. (c) The ratio of β-actin-GFP fluorescence intensity at bleached stereocilia tips was compared with that of unbleached stereocilia tips (20 or more cells analysed per time point). The fraction recovered was plotted and fitted with a single exponential curve (T1/2=215 min). Error bars represent s.d., *P<0.01, analysis of variance. Scale bars, 5 μm.

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