Figure 2.

The internalization and not the auxin inhibition of internalization of the endogenous PIN2 is impaired in gsnor1-3 mutant. The immunolocalizations were performed using PIN2-specific antibodies (Wang et al., 2013) and Cy3-labeled anti-rabbit secondary antibodies (Sigma-Aldrich) on the primary roots of 6-day-old seedlings treated with Mock (A,D), 50 μM BFA for 60 min (B,E) or with 10 μM 2, 4-D for 30 min, followed by treatment with 50 μM BFA+ 10 μM 2, 4-D for additional 60 min (C,F). Images were captured using CLSM (Leica TCS SP5 AOBS). The numbers of BFA bodies (see white arrows) were counted and the fluorescence intensities (FI) at the BFA bodies and at the PM were measured, respectively, using Image J (http://rsb.info.nih.gov/ij) and the statistical data were summarized (G–J). (G) Fluorescence intensity (FI) at the PM; (H) Number of BFA bodies per cell; (I) Relative fluorescence intensities (FI) of the BFA bodies; (J) The fluorescence intensity (FI) ratios of the BFA bodies/PM. Bar = 50 μm. ^***Indicates significant differences between Col-0 and gsnor1-3 by Student's t-test at 0.001 level.