Extended Data Fig. 8: KOIN interacts with 14-3-3λ and modulates of CBF signaling under cold stress.
From: A receptor–kinase cascade confers cold-induced root growth inhibition in Arabidopsis
a, Co-IP assays in Arabidopsis protoplasts showing interaction between KOIN-GFP and Flag-HA-14-3-3λ. Proteins were detected using anti-GFP and anti-HA antibodies. b, In vitro pull-down assay showing the interaction between KOIN and 14-3-3λ. Anti-His antibodies were used for detection. c, BiFC assay in Arabidopsis protoplasts showing KOIN-14-3-3λ interaction at the plasma membrane. FM4-64 staining was used to visualize endosomes. BRI1–YFPN+14-3-3λ–YFPC served as a negative control. Negative control (BRI1-YFPN+14-3-3λ-YFPC) is shown. Scale bar: 5 μm. d,e, Transcript levels of CBFs (d) and their target genes (e) in Col-0, koin-2, koin-2 KOIN complementation lines under cold treatment. Expression is shown relative to untreated Col-0 (set as 1.0), with ACTIN2/8 as internal reference. Data represent mean ± s.d. (n = 3, one biological replicate), representative of three independent experiments. **P < 0.01 (one-way ANOVA with Tukey’s test). Exact P values are provided in the Supplementary Table 2.
