Fig. 7: Phosphorylation at the main CPK1-target sites is required for the activation of CNGC5/6/9 in HEK293T cells. | Nature Communications

Fig. 7: Phosphorylation at the main CPK1-target sites is required for the activation of CNGC5/6/9 in HEK293T cells.

From: CPK1 activates CNGCs through phosphorylation for Ca2+ signaling to promote root hair growth in Arabidopsis

Fig. 7

CPK1, the wild type CNGC5/6/9, and iCNGCs with a S-to-A point mutation were cloned into the vector pCI-neo-IRES-YC3.6, and were then expressed in HEK293T cells individually or in a combination as indicated. HEK293T cells with bright YC3.6 fluorescent signal were used for cytosolic Ca2+ imaging and patch clamping analysis. a–c Cytosolic Ca2+ imaging data show the strong inhibition of CNGC5 (a), CNGC6 (b), and CNGC9 (c) by the S-to-A point mutations at the main CPK1-target sites compared to the strong activation of wild type CNGCs by CPK1. d–i Patch clamping data show the inhibition of CNGC5 (d, e), CNGC6 (f, g), and CNGC9 (h, i) by the S-to-A point mutations at the main CPK1-target sites compared to the activation of wild type CNGC5/6/9 by CPK1. C5S20A, C6S27A, and C9S26A represent CNGC5S20A, CNGC6S27A, and CNGC9S26A, respectively. The numbers of HEK293T cells tested are indicated in brackets, and data are presented as means ± SEM in (a–c), (e), (g), and (i). Source data are provided as a Source Data file.

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