Extended Data Fig. 8: CCA1 interacts with MPK1/2/7/14 in yeast two-hybrid and split luciferase complementation imaging assays.

a-d Y2H assay showing CCA1 interaction with MPK1 (a), MPK2 (b), MPK7 (c) and MPK14 (d). Co-transformation of CCA1-BD with empty AD or AD-MPK1/2/7/14, or of AD-MPK1/2/7/14 with empty BD vector. AD and BD vectors served as negative controls. SD/-Leu-Trp, synthetic defined (SD) medium lacking Leu and Trp; SD/-Leu-Trp-His-Ade, SD medium lacking Leu, Trp, His, and Ade. e-h Split-LUC complementation imaging assay in N. benthamiana testing interaction between CCA1 and MPK1 (e), MPK2 (f), MPK7 (g) and MPK14 (h). Indicated constructs were transiently expressed in N. benthamiana by Agrobacterium-mediated transformation, and the GUS (β-glucuronidase) protein served as negative controls. nLUC, N-terminal fragment of firefly luciferase; cLUC, C-terminal fragment of firefly luciferase.