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. 2014 Mar 21;26(3):1166–1182. doi: 10.1105/tpc.113.117069

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© 2014 American Society of Plant Biologists

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Figure 6. — Ser-294 in SOS2 Is Required for the Repression of SOS2 by 14-3-3 λ.

(A) Analysis of the phosphorylation status of SOS2^K40N, SOS2^K40N-S294A, and SOS2^K40N-S294D in Arabidopsis leaf protoplasts. The phosphorylation status of transiently expressed Myc-tagged SOS2^K40N, SOS2^K40N-S294A, and SOS2^K40N-S294D (shown in Figure 5G) was analyzed using anti-phosphoserine/phosphothreonine (anti-p-S/T) polyclonal antibodies. IB, immunoblot. Experimental details are provided in Methods.

(B) Kinase assays for recombinant SOS2, SOS2^S294A, and SOS2^S294D with or without 14-3-3 λ. SCaBP8 (SC8) was used as the substrate. CBB, Coomassie Brilliant Blue; Autorad, autoradiograph. Experimental details are provided in Methods.

(C) Kinase assay for Myc-SOS2, -SOS2^S294A, and -SOS2^S294D purified from Col-0 with or without 14-3-3 λ. SCaBP8 was used as the substrate. Experimental details are provided in Methods.

(D) Kinase assay for Myc-SOS2, -SOS2^S294A, and -SOS2^S294D purified from the 14-3-3 λ κ double mutant. SCaBP8 was used as the substrate. Experimental details are provided in Methods.