Fig. 3: RLP4 responds to changes in cell wall mechanical status.
From: A self-regulatory cell-wall-sensing module at cell edges controls plant growth
a,b, CLSM maximum-intensity projections of lateral root epidermal meristems co-expressing pUBQ10::RLP4–RFP and pRAB-A5c::YFP–RAB-A5c after three days of treatment with 2.5 nM IXB or DMSO. c, Manders’s colocalization coefficients between RLP4s–RFP and various membrane markers with or without IXB as shown in a,b. N = 14 regions from two roots (VHA-a1–GFP DMSO and YFP–RAB-A5c DMSO), 15 regions from two roots (VHA-A1–GFP IXB), 16 regions from two roots (YFP–RAB-A5c IXB) or 17 regions from three roots (the remaining combinations). *P < 0.05; ***P < 0.001 (ANOVA with post-hoc Tukey test). See Methods for an explanation of the box plots. d–f, CLSM maximum-intensity projections of a lateral root epidermal meristem coexpressing RLP4–RFP and YFP–NPSN12 before and immediately after cell ablation (asterisks). g, Fluorescence intensity ratio of RLP4–RFP and YFP–NPSN12 after and before ablation on transverse midplane sections of epidermal meristem cells undergoing deformation after ablation like those shown in d–f. The lines indicate average values (N ≥ 62 edges from five roots), and the shaded areas show ±1 s.d. The asterisks indicate significant differences between RLP4–RFP and YFP–NPSN12 (one-way ANOVA and post-hoc Tukey test; ***P < 0.001). h–k, XZ projections showing the same anticlinal cell edges of lateral roots co-expressing pUBQ10::RLP4–RFP, YFP–NPSN12 and β-oestradiol-inducible A-L2 after 16 h of treatment with DMSO (h,i) or 10 µM β-oestradiol (j,k) before (h,j) and after (i,k) microneedle ablation. l, Fluorescence intensity of RLP4–RFP before and after ablation on midplane sections of epidermal meristem cells like those in h–k. The lines indicate average fluorescence intensity (N = 24 (DMSO) and N = 32 (β-oestradiol) edges from three roots, respectively), and the shaded areas show ±1 s.d. The asterisks indicate significant differences in RLP4–RFP intensity between DMSO and β-oestradiol treatments (one-way ANOVA and post-hoc Tukey test; ***P < 0.001). Before the ablation, there was no significant difference in RLP4–RFP intensity between treatments. m–p, CLSM maximum-intensity projections of lateral root epidermal meristem cells coexpressing RLP4–RFP and YFP–RAB-A5c in the absence (m,n) or presence (o,p) of inducible A-L2 before (m,o) and immediately after cell ablation (n,p; asterisks). Note that RLP4–RFP and YFP–RAB-A5c show increased retention at the cell edge in the presence of A-L2 (arrows). Scale bars, 10 µm (d–f), 5 µm (a,b,m–p) or 2 µm (h–k).
