DEV004788 Supplementary Material

Files in this Data Supplement:

• Supplemental Table S1 -

  Adobe PDF

• Supplemental Figure S1 -

  Fig. S1. Floral phenotypes of the er-family. (A,D) Wild type; (B,E) er-105; (C,F) er-105 erl1-2 erl2-1/+. Flowers of the er-105 erl1-2 erl2-1/+ mutant do not exhibit floral patterning defects.

• Supplemental Figure S2 -

  Fig. S2. DAPI staining of mature ovules. Representative images of mature ovules of (A) wild type (WT) and (B) er-105 erl1-2 erl2-1/+ and (C) of a wild-type root tip stained with DAPI. Root tip is shown for a 2N nuclei control. Ovules were removed from siliques using forceps and mounted on slides in 1 μg/ml DAPI solution. Roots were mounted directly on slides in 1 μg/ml DAPI solution. DAPI fluorescence was viewed using a Radiance 2000MP confocal microscope (Biorad, Hercules, CA) and merged with the DIC image using Photoshop (Adobe, San Jose, CA). No qualitative difference in the size (area) of the nuclei between wild type and er-105 erl1-2 erl2-1/+ was observed, suggesting er-105 erl1-2 erl2-1/+ integument cells do not undergo endoreduplication.

• Supplemental Figure S3 -

  Fig. S3. RT-PCR analysis of CDKB and cyclin A2 genes in stage 12 carpels. Loss of ER-family function misregulates core cell-cycle regulators. ACTIN serves as a positive control. Real-time PCR was performed on CYCLINA2;2 and CYCLINA2;3 (see Fig. 4).

• Supplemental Figure S4 -

  Fig. S4. ERECTA and ERL1 in situ probes do not cross-hybridize. ERL1 and ER antisense probe hybridization in er-family mutant backgrounds. ER antisense probe in wild type (A) and er-105 (B). ERL1 antisense probe in wild type (C), erl1-2 erl2-1 (D) and er-105 erl1-2 erl2-1/+ (E). Hybridization between family members was not detected using the conditions and probes in this study.

• Supplemental Figure S5 -

  Fig. S5. Comparison of ovule development in er-105 erl1-2 erl2-1/+ and er-105 erl1-2 erl2-1. (A) er-105 erl1-2 erl2-1/+ and (B) er-105 erl1-2 erl2-1.The complete absence of ER-family signaling results in severely stunted ovules, with almost no extension of the funiculus or integuments. Note that overall ovule patterning is not disrupted. n, nucellus; ii, inner intergument; oi, outer integument; f, funiculus.