Figure 5: Suppression of the ros1 epidermal patterning phenotype by RdDM mutations. | Nature Communications

Figure 5: Suppression of the ros1 epidermal patterning phenotype by RdDM mutations.

From: Overproduction of stomatal lineage cells in Arabidopsis mutants defective in active DNA demethylation

Figure 5

(a–f) Adaxial epidermal cell phenotype of 3-day-old seedlings. (a) nrpd1-3, (b) nrpd1-3ros1-4, (c) hda6, (d) hda6ros1-4, (e) drm1-2drm2-2cmt3-13 (ddc) and (f) drm1-2drm2-2cmt3-13ros1-4 (ddcros1-4). Images in a–f are at the same magnification. Scale bar, 30 μm. (g) Numbers of clustered small cells and stomata for Col, ros1-4, nrpd1-3, nrpd1-3ros1-4, hda6, hda6ros1-4, drm1drm2cmt3 (ddc), drm1drm2cmt3ros1-4 (ddcros1-4). Values are means+s.d.’s per 25,000 μm2 of 3-day-old cotyledon adaxial epidermis (n=3). Student’s t-test, *P<0.01 (significantly different from Col). (h) Bisulfite sequencing analysis of the EPF2 promoter, and (i) quantitative RT–PCR assay of EPF2 transcript levels in Col, ros1-4, nrpd1-3, nrpd1-3ros1-4, hda6 and hda6ros1-4, drm1drm2cmt3 (ddc), drm1drm2cmt3ros1-4 (ddcros1-4). Student’s t-test, *P<0.01 (significantly different from ros1-4). Error bars represent s.d.’s (n=4). (j) Small RNA northern blot analysis in Col, nrpd1-3, ros1-4 and nrpd1-3ros1-4. tRNA and other RNA bands stained with ethidium bromide (EtBr) were used as loading control. miR167 was detected as an internal control. Arrows and arrowheads indicate the positions of size markers, 21 nt and 24 nt, respectively.

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