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. 2009 May 20;96(10):4063–4074. doi: 10.1016/j.bpj.2009.02.055

Figure 2.

Figure 2

KDC1 is targeted to the membrane only in tandem with KAT1. Confocal laser-scanning microscopical analyses on oocytes expressing KAT1-EGFP (A), KAT1/KDC1-EGFP (B), and KDC1-EGFP (C) microinjected with Fura Red. The left column shows the fluorescence collected from an x-y plane of the oocyte. Panels A and B show a green layer (belonging to GFP signal) external to the red one (belonging to Fura Red signal); the yellow layer is due to the colocalization of the two fluorophores. In panel C there is no green layer. The right column shows the results obtained by a spectral unmixing analysis, applied on every injected oocyte: traces show normalized fluorescence intensity for a particular region of interest (ROI) of the two fluorophores as a function of the distance z (0 coincides with the slide plane and 40 μm with the most internal plane of the oocyte reached). The distance between the two peaks, d(EGFP-Fura Red), is significant in the case of KAT1-EGFP (2.6 ± 0.3 μm) and KAT1/KDC1-EGFP (2.5 ± 0.3 μm), whereas for KDC1-EGFP the two peaks practically coincide (−0.5 ± 0.3 μm). Data are mean values ± SE from more than 20 ROI.