Figure 6.

AKINβ2 is detected in different protein complexes with SnRK α-subunits AKIN10 and AKIN11 in vivo. Arabidopsis cell suspensions were transformed with Agrobacterium strains to express myc–AKIN10, AKIN11–myc and HA–AKINβ2 separately. In co-transformation experiments, HA–AKINβ2 was expressed together with either myc–AKIN10 or AKIN11–myc. (A) Immunoblotting of protein extracts (50 µg) from cell lines co-expressing HA–AKINβ2 with c-Myc-tagged SnRK α-subunits using anti-c-Myc and anti-HA antibodies. (B) From each single and double transformed cell line 1 mg protein extract was subjected to immunoaffinity purification on either an anti-HA IgG or anti-c-Myc IgG matrix. Proteins eluted from the anti-HA IgG and anti-c-Myc IgG matrices using HA and c-Myc peptides, respectively, were immunoblotted and detected with anti-HA and anti-c-Myc antibodies. The symbols + and – indicate the presence or absence of constructs in the transformation assays. IP and ID refer to the antibodies used for immunoprecipitation (IP) and immunodetection (ID), respectively.