Fig. 3: Sucrose induces the KIN10 protein accumulation.

a Immunoblot analysis of the protein levels of KIN10-myc in pKIN10::KIN10-myc transgenic plants using anti-myc antibody. Seedlings of pKIN10::KIN10-myc transgenic plants were grown in sugar free ½ MS liquid medium for 3 days and then treated with 1% sucrose or 1% mannitol for different time periods. b Quantitative RT-PCR analysis the expression of KIN10 in response to sucrose. Wild-type seedlings were grown in sugar free 1/2 MS liquid medium for 3 days then treated with 1% sucrose for different times. PP2A gene was analyzed as an internal control. Error bars represent standard deviation of four independent experiments. Different letters above the bars indicated statistically significant differences between the samples (ANOVA analysis followed by Uncorrected Fisher’s LSD multiple comparisons test, p < 0.05). c Sucrose induces KIN10 protein accumulation in p35S::KIN10-myc transgenic plants. Seedlings of p35S::KIN10-myc transgenic plants were grown in sugar free ½ MS liquid medium for 3 days and then treated with 1% sucrose or 1% mannitol for different time periods. d Immunoblot analysis the effects of cycloheximide (CHX) on the Sucrose-induced the KIN10 protein accumulation. Seedlings of p35S::KIN10-myc transgenic plants were grown in sugar free ½ MS liquid medium for 3 days and then treated with or without 1% sucrose and 50 μM CHX for different time periods. e Quantification analysis of CHX effect on sucrose-induced KIN10 protein accumulation. The ratio of immunoprecipitated KIN10-myc to actin was quantified by ImageJ software. Values for different time points after sucrose addition with or without CHX treatment are given as mean ± S.E. (n = 3). Asterisk above dots indicates significant inhibition of KIN10 protein accumulation by CHX treatment at different time points (ANOVA analysis followed by Uncorrected Fisher’s LSD multiple comparisons test, *p < 0.05; **p < 0.01; ****p < 0.0001).