SlCipk6 and SlRd2 interact in planta and the complex localizes in the cytosol. A, Protein extracts from N. benthamiana leaves agro-infiltrated with GFP-SlRd2 and SlCipk6(T172D)-Myc, which is fused to two copies of the protein A IgG binding domains (2xIgG-BD), were incubated with IgG agarose beads, eluted and analyzed by immunoblotting using anti-Myc for SlCipk6 and anti-GFP for SlRd2. GFP protein was used as a negative control. No interaction was detected when SlCipk6(T172D) was agro-infiltrated with GFP (right upper panel). B, Fluorescence images of N. benthamiana leaf sections agroinfiltrated with SlRd2-YFPC and SlCipk6-YFPN. Colocalization analyses of SlCipk6/SlRd2 complexes (yellow, a and d) with the fluorescent membrane marker dye FM4-64 (red, b) and the cytoplasmic marker AtFKBP12-CFP (cyan, e; Faure et al., 1998). Merged images are shown in (c) and (f), respectively. White arrows show the cytoplasmic localization observed for the SlCipk6/SlRd2 complexes. Localization analysis of YFPN (g) and YFPC (h) empty vectors and SlCbl10/SlRd2 (i) were used as an experiment controls. Bars = 10 μm.