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. 2020 Feb 6;180(3):440–453.e18. doi: 10.1016/j.cell.2020.01.013

Figure S2.

Figure S2

Laser Ablation-induced MAMP Responsiveness Rely on Cell Damage Extent and Functional FLS2, Related to Figure 2

(A and B) Representative images (A) and quantitative analysis by column scatterplot (B) of PER5 responsiveness after laser ablation of different cell types in differentiated roots. Laser ablation and all images were at 25 endodermal cells after the onset of cell elongation. Nuclear-localized mVENUS signals for each PRR reporter (green) co-visualized with plasma membrane marker, pUBQ10::RCIA2A-tdTomato (red). Maximum projections of Z stack of mVENUS signals were combined with single red-channel images (see Figure 2A). White asterisks indicate laser-ablated cells. Arrows indicate PER5 responsive nuclei. Scale bar, 50 μm. Each circle in (B) represents individual laser ablation event of one root (n = 12 roots). Graph depicts mean values and SD (error bars). Asterisks indicate significant differences between means by ANOVA and Tuckey’s test (p < 0.001). ns, not significant. Ep, epidermis; Co, cortex; En, endodermis; St, stele. Scale bar, 50 μm.

(C and D) Representative images (C) and quantification by column scatterplot (D) of MAMP responsiveness after laser ablation of different epidermal cell numbers with or without flg22 for 6 h in differentiated roots. Nuclear-localized mVENUS signals of FRK1 and PER5 reporters (green) co-visualized with the plasma membrane marker, pUBQ10::RCI2A:tdTomato (red). White asterisk indicates damaged cell by laser ablation. Scale bar, 50 μm. Each circle in (D) represents individual laser ablation event (n = 12). Data represent mean values and SD (error bars). 1 epi, one epidermal cell; 2 epi, two epidermal cells; etc.

(E and F) Orthogonal views (E) and RID quantification (F) of FRK1 and PER5 responsiveness in WT and fls2 mutant background after combining without (-ablation) or with (+ablation) damage of epidermal cells in the absence or presence of flg22 for 6 h. Scale bar, 50 μm. Boxplot centers in (F) show median. Asterisks indicate significant differences between means (p < 0.001) by ANOVA and Tukey’s test analysis (n = 10 independent ablation events). ns, not significant. RID, see legend Figure 1C.