Figure 2.

PI4Kγ2 interacts with E3 ligase MIEL1. A, Yeast two-hybrid assays revealed a direct interaction between PI4Kγ2 and MIEL1. Transformed yeast cells were grown on SD-Leu-Trp or SD-Leu-Trp-His-Ade. AD, Activation domain; BD, binding domain. B, Observation of transiently expressed fusion proteins in Arabidopsis protoplasts showed that PI4Kγ2 colocalizes with MIEL1. Scale bars = 10 μm. C, BiFC assays confirmed the MIEL1-PI4Kγ2 interaction in planta. A model no. FV1000 confocal microscope (Olympus) was used and collection wavelength was set at 510 to 550 nm to strictly filter out the autofluorescence (details in “Materials and Methods” section), resulting in two control images (upper, middle and right) that are blank. Scale bars = 30 μm.