Fig. 3: N-glycosylation positively controls SnRK2.2/2.3’s redistribution from the nucleus to the peroxisomes.
From: N-glycosylation of SnRK2s affects NADPH maintenance in peroxisomes during prolonged ABA signalling
a Subcellular dynamics of SnRK2.2-GFP and SnRK2.3-GFP under snrk2.2/2.3 background during prolonged ABA signalling. 4-day-old seedlings of report lines were treated with 50 μM ABA for different time periods. b Quantification of SnRK2.2-GFP and SnRK2.3-GFP fluorescence signal shown in (a). The red dashed line represents the number of peroxisomes containing SnRK2.2-GFP or SnRK2.3-GFP in one cell. The blue line represents the fluorescence intensity in the nucleus in one cell. n = 10 cells. “Pero” means peroxisome. c Overlap between SnRK2.2/2.3-GFP and peroxisomal membrane mark gene, dsRed-mPTSPEX26. Photographs were taken after 50 μM ABA treatment for 48 h. “n” means nucleus, “c” means cytoplasm and “pero” means peroxisome. Scale bars, 5 μm. n = 8 Arabidopsis roots. d STT3A positively controls SnRK2.2-GFP’s redistribution from the nucleus to the peroxisomes during prolonged ABA signalling. SnRK2.2-GFP report line was crossed into backgrounds of Col-0, stt3a-2, and STT3A-OE, respectively. The fluorescent signal is recorded after treatment with 50 μM ABA for different time periods. Scale bars, 5 μm. e Quantification of the number of peroxisomes containing SnRK2.2-GFP per cell shown in (d). n = 16 cells. “Pero” means peroxisome. f N-glycosylation of SnRK2.3 positively controls its redistribution. 4-day-seedlings of SnRK2.3-GFP and SnRK2.3N323A-GFP transgenic plants were treated with 50 μM ABA for different time periods. Scale bars, 5 μm. g Quantification of GFP labelled peroxisomes per cell. n = 9 cells. The blue and red lines represent SnRK2.3-GFP and SnRK2.3N323A-GFP, respectively. “Pero” means peroxisome. Data are presented as means ± s.d. h Quantification of fluorescent intensity in the nucleus in each cell shown in (f). n = 9-13 cells. i, j The expression levels of RAB18 and NCED3. The transgenic plants were treated with 50 μM ABA for different time periods. 3 biological replicates for each time point. Data are presented as means ± s.d., and asterisks indicate significant differences (*P-value < 0.05, **P-value < 0.01, ***P-value < 0.001), as determined by two-tailed paired t tests. “ns” means no significant difference.
