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Files in this Data Supplement:
Fig. S1. GUS reporter analysis of HEC expression. (A-C) GUS expression in the HEC1p::HEC1:GUS reporter line. Transverse section of a stage 8 flower (A), a longitudinal section of a stage 9-10 flower (B) and a transverse section of a stage 12 gynoecium (C). HEC1:GUS expression duplicates the pattern seen by in situ analysis for septum and stigma, but not for ovule funiculus. (D) Longitudinal section of an early stage 12 flower. HEC2p::GUS reporter line duplicates the septum and stigma expression pattern seen by in situ analysis. (E) Transverse section of a stage 14 gynoecium. HEC3p::GUS reporter line shows that HEC3 funiculus expression persists even after pollination (arrowheads), whereas septum and transmitting tract expression are absent. There is also evidence of vascular expression (arrow). Scale bars: 50 μm.
Fig. S2. RT-PCR analysis of T-DNA mutants and HEC2-RNAi lines. (A) RT-PCR showing that a full-length HEC1 transcript is not produced in the hec1 mutant. In order to determine how much of the PCR product was the result of DNA contamination, a control reaction without reverse transcriptase (RT) was performed. TUBULIN was used as a loading control. (B) RT-PCR showing that a full-length HEC3 transcript is not produced in the hec3 mutant. A control reaction without RT was performed. TUBULIN was used as a loading control. (C) RT-PCR showing that reduced levels of HEC2 transcript are produced in two independent HEC2-RNAi hec1 hec3 lines relative to wild type. A control reaction without RT was performed. Levels of the SPT transcript are unaffected. TUBULIN was used as a loading control. (D) RT-PCR analysis of HEC2, HEC1 and HEC3 transcript levels in four independent HEC2-RNAi lines in the wild-type (Col-0) background. HEC2 and HEC1 RNA levels are reduced to about the same degree. HEC3 RNA levels are slightly reduced. TUBULIN was used as a loading control. To determine how much of the HEC2 PCR product was the result of DNA contamination, a control reaction without RT was performed.
Fig. S3. Defects in early septum development in the hec1 hec3 double mutant. (A,C) Transverse sections through early stage 12 gynoecia stained with Alcian Blue and Fast Red. (B,D) Transverse sections through late stage 12 gynoecia stained with Alcian Blue and Fast Red. More darkly staining ECM is just starting to be visible early in stage 12 (arrowheads) in both wild type (A) and hec1 hec3 (C) gynoecia. However, hec1 hec3 shows less staining and a narrower septum. By late stage 12 in wild-type (B), transmitting tract cells are secreting blue-staining ECM (arrowheads), but the hec1 hec3 transmitting tract (D) is narrower and is producing less ECM. Scale bars: 50 μm.
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