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. Author manuscript; available in PMC: 2015 Dec 25.
Published in final edited form as: Nature. 2015 Jun 17;522(7557):439–443. doi: 10.1038/nature14561

Extended Data Fig. 3. STOMAGEN overexpression promotes stomatal differentiation in genetic backgrounds missing/blocking EPF2-ER and EPF1-ERL1 signaling components.

Extended Data Fig. 3

(a-j) Representative confocal images of cotyledon abaxial epidermis from 10-day-old light-grown transgenic seedlings of the following genotypes, each carrying Est∷STOMAGEN construct: epf2 (a, b); Dominant-negative ER (ERΔK) in er (c, d); epf1 (e, f); Dominant-negative ERL1 (ERLΔK) in erl1 (g, h); er erl1 erl2 (i, j). For each genotype, a control uninduced phenotypes (a, c, e, g, i) and induced STOMAGEN overexpression (iSTOMAGEN) (b, d, f, h, j) are shown. Blocking ER or lacking EPF2 produces small stomatal-lineage cells due to excessive entry divisions (a and c; brackets). iSTOMAGEN confers stomatal clusters and small stomatal-lineage cells are no longer present (b and d). Blocking ERL1 or lacking EPF1 causes a stomatal pairing due to a violation of one-cell-spacing rule (e and g; dots). iSTOMAGEN enhances stomatal cluster phenotype in these genotypes (f and h). iSTOMAGEN does not enhance stomatal clustering defects in er erl1 erl2 (i and j). Images were taken under the same magnification. Scale bars = 30 μm. n=29 (a); n=24 (b); n=16 (c); n=17 (d); n=22 (e); n=23 (f); n=17 (g); n=20 (h); n=24 (i); n=24 (j). (k-m) Stomatal Density (SD: number of stomata per mm2) (k); Stomatal Index (SI: % of number of stomata per stomata+ non-stomatal epidermal cells) (l); and Stomatal Cluster Distribution (in %)(m) from 10-day-old abaxial cotyledons of transgenic lines of each genotype carrying Est∷STOMAGEN construct. -, no induction; +, induced by 10 μM estradiol. Stomagen overexpression significantly increases SD in all genotypes except for er erl1 erl2 and tmm. Error bars, S.E.M. ***, p< 0.001; **, p<0.01; NS, not significant; Welch 2-sample T test. Number of seedlings subjected to analysis, n=14-16. Total numbers of stomata counted: wt, no induction, 1277, induction, 2639; epf1 no induction, 1390, induction 3485; ERL1ΔK erl1, no induction 1573, induction, 3991; epf2, no induction, 2502, induction, 3317; ERΔKer, no induction, 2899, induction, 4397; tmm, no induction, 2948, induction, 3212; er erl1 erl2, no induction, 4454, induction, 4464. All genotypes carry Est∷STOMAGEN. wt, no induction, n=16, induction, n=14; epf1 no induction, n=16, induction n=17; ERL1ΔK erl1, no induction n=15, induction, n=15; epf2, no induction, n=15, induction, n=15; ERΔK er, no induction, n=15, induction, n=15; tmm, no induction, n=15, induction, n=15; er erl1 erl2, no induction, n=15, induction, n=15.