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. 2013 May 22;8(5):e64096. doi: 10.1371/journal.pone.0064096

Figure 6. Role of the di-leucine motif in the localization of GIRK5.

Figure 6

A) Confocal microscopy images of EGFP chimeras. Removal of the hydrophobic leucine and isoleucine residues in the nonphosphorylated GIRK5 (YLI/AAA), targeted the channel equally to both poles. Remarkably, alanine mutation of the whole di-leucine sorting signal (YELI/AAAA) produced GIRK5 polarization to the animal pole. Scale bar: 250 µm. B) Fluorescence quantification. Error bars correspond to mean ± SD, n = 4–6. Triangle indicates significant differences of oocytes compared to Y16A (P<0.001; One-Way ANOVA). Asterisk indicates significant differences of oocytes compared between them (P>0.001; One-Way ANOVA). The statistic significance between samples is the same for the animal and vegetal pole. C) Immunoblot analysis revealed bands that correspond to the expected weight of 75 kDa of the EGFP-GIRK5 constructs: 1) Non-injected, 2) GIRK5-Δ25, 3) GIRK5-WT, 4) GIRK5-Y/A, 5) GIRK5-YLI/AAA and 6) GIRK5-YELI/AAAA.