Fig. 3: PRAF proteins are localized to the plasma membrane and endomembrane. | Nature Communications

Fig. 3: PRAF proteins are localized to the plasma membrane and endomembrane.

From: Connected function of PRAF/RLD and GNOM in membrane trafficking controls intrinsic cell polarity in plants

Fig. 3: PRAF proteins are localized to the plasma membrane and endomembrane.The alternative text for this image may have been generated using AI.

a YFP-tagged PRAF8 rescues growth defects of praf5c;8c;9c. Comparison of 4-week-old plants of the wild type, praf5c;8c;9c, and PRAF8p::PRAF8-YFP in praf5c;8c;9c. b–c Protein localization of PRAF8 (green) at the native level (b) or overexpressed in the stomatal lineage cells (PRAF8++, c) in 4-day-old adaxial side cotyledon epidermis. Inset in (b) shows an enlarged view of guard cells exhibiting endosomal localization of PRAF8 (arrowheads). d–e PRAF8 polarization (arrows) requires BASL. Overexpressed PRAF8 (TMMp::Venus-PRAF8, green) in wild type (d) and in basl-2 (e), respectively. Data represent the results of three independent experiments. Magenta, PI-stained cell outlines. f Quantification of PRAF8 polarization in (d, e). Box plots show first and third quartile (box), median (line) and mean (cross). n, # stomatal lineage cells. Student’s unpaired t tests were used. Two-sided P value is 0.0001. ***P < 0.001. g Confocal images show individual protein expression and co-expression of TMMp::mCherry-PRAF8 (magenta) with BASLp::GFP-BASL (green). h Left, confocal image shows protein localization of PRAF8p::myr-PRAF8-YFP (green) in 4-day-old adaxial side cotyledon epidermis. Inset, enlarged view of guard cells. Note the reduced association of myr-PRAF8-YFP with intracellular particles compared to the wild-type PRAF8-YFP (inset in (b)). Right, quantification of # intracellular puncta for PRAF8p::PRAF8-YFP; praf5c;8c;9c and for PRAF8p::myr-PRAF8-YFP; praf5c;8c;9c, respectively. Box plots show first and third quartile (box), median (line) and mean (cross). n, # guard cell (GC) pairs counted. Student’s unpaired t tests were used. Two-sided P value is 0.0005. ***P < 0.001. i Expression of myr-PRAF8-YFP partially rescued the growth defects of praf5c;8c;9c mutants. Plants were about 4-week old. (z) stands for z-stacked confocal images. Scale bars in (a) and (i) are 1 cm and all others are 10 μm. Data represent results of three independent experiments.

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