Figure 8.

eAna2(12D/E)-mNG is not recruited to centrioles efficiently. (A i) Images show the typical centriolar recruitment dynamics of WT Ana2-mNG or eAna2(12D/E)-mNG in an embryo also expressing one copy of the endogenous untagged ana2 gene during nuclear cycle 12—aligned to nuclear envelope breakdown (NEB; t = 0). Images were obtained by superimposing all the centrioles at each time point and averaging their fluorescence (scale bar = 1 µm). Note that the centrioles in the embryo expressing eAna2(12D/E)-mNG were very dim so their brightness has been enhanced by 2X relative to the WT control. (ii) Graph shows normalized (mean ± SEM) centriolar recruitment dynamics of either WT Ana2-mNG (black) or eAna2(12D/E)-mNG (blue) expressed in the presence of 1 copy of the endogenous untagged ana2 gene during nuclear cycle 12. Data were aligned to nuclear envelope breakdown (NEB; t = 0). N ≥ 11 embryos, n ∼ 100–150 centriole pairs per embryo. (B) Bar charts quantify the normalised initial and maximal fluorescence intensity (mean ± SEM). Each data point represents the average value of all centrioles measured in an individual embryo. Statistical significance was assessed using an unpaired t test with Welch’s correction (****, P < 0.0001).