Subcellular localization of predicted plant KASH proteins. (A and C) GFP-tagged SINE1, SINE2, SINE3, SINE4, and SINE5 under control of the 35S promoter were stably expressed in wild type (A) or sun1-KO sun2-KD (C), respectively. (B) GFP-tagged SINE1ΔTVPT, SINE2ΔLVPT, SINE3ΔPLPT, SINE4ΔLVPT, and SINE5ΔLVPT driven by the 35S promoter were stably expressed in wild-type A. thaliana. Root tip cells were imaged using confocal microscopy. Bars, 5 µm. GFP signal is shown in green. Images in the second column of A are overlays of GFP and transmitted light images. Cell-to-cell variability of GFP fusion protein abundance was seen in all images. (D) The NLI was calculated and compared. As illustrated in the top of D, two maximum NE intensities (N1 and N2) and two maximum cytoplasmic intensities (C1 and C2) along a random line across a cell were chosen to calculate the NLI, which equals (N1 + N2)/(C1 + C2). Asterisks in D represent significant statistic differences when compared with GFP-tagged wild-type SINEs in wild-type A. thaliana (P < 0.01, two-tailed t test, n = 30). Error bars show SEMs.
