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Additional file 1: Figure S1. of The Mediator complex subunits MED25/PFT1 and MED8 are required for transcriptional responses to changes in cell wall arabinose composition and glucose treatment in Arabidopsis thaliana

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posted on 2015-09-05, 05:00 authored by Mathilde Seguela-Arnaud, Caroline Smith, Marcos Uribe, Sean May, Harry Fischl, Neil McKenzie, Michael Bevan
Scatter plots of Principle Components 1, 2 and 3 identified from FTIR measurements of cell wall composition. Figure S2. JA responsive genes are not up- regulated in hsr8. Quantitative Real-time PCR analysis of VSP1, VSP2 and ERF1 mRNA levels in Col and hsr8. Seedlings were grown vertically in the dark for 14 days on MS medium in the presence of 1 % glucose. Errors bars represent SD from three biological replicates. **, p < 0.01 comparing Col to hsr8-1 (Student’s t- test). Figure S3. Sugar- regulated gene expression in the single mutant pft1-2. (A) to (E) Quantitative Real-time PCR analysis of mRNA levels of the glucose-responsive genes APL3, BAM, GBSS1, GPT2, PDC1 in Col and pft1-2 in response to glucose. Seedlings were grown on MS medium supplemented with 0.5 % glucose in constant light. After 7 days, the seedlings were transferred for 24 h to glucose-free MS liquid medium (solid bars) and then treated for 6 h with 3 % glucose (dashed bars). Errors bars represent SD from three biological replicates. Data shown is representative of three independent experiments. **, p < 0.01 comparing glucose responses in Col to pft1-2 (Student’s t- test). (G) and (H) Quantitative Real-time PCR analysis of mRNA levels of the anthocyanin biosynthesis genes CHS, TT6 and FLS in Col and pft1-2 in response to glucose. Seedlings were grown on MS medium supplemented with 0.5 % glucose in constant light. After 7 days, the seedlings were transferred for 24 h to glucose-free MS liquid medium (solid bars) and then treated for 6 h with 3 % glucose (dashed bars). Errors bars represent SD from three biological replicates. Data shown is representative of three independent experiments. **, p < 0.01 comparing glucose responses in Col to pft1-2 (Student’s t- test). Relative transcript levels (RTL) were calculated relative to the transcript level of the reference gene TUB6 (At5g12250). (I) Anthocyanin accumulation in response to glucose in Col and pft1-2 in response to glucose. Seedlings were grown in continuous light for 7 days on MS containing 1 % glucose (solid bars) or 3 % glucose (dashed bars). Errors bars represent SD from three biological replicates. **, p < 0.01 comparing glucose responses in Col to pft1-2 (Student’s t- test). Data shown is representative of two independent experiments. Figure S4. PFT1 regulates the expression of genes encoding cell wall- related genes. Quantitative Real-time PCR analysis of genes encoding the pectin methylesterases PME17 (A), PME41 (B), and the extensin proteins AtEXT3 (C) and AtEXT4 (D) in Col, hsr8-1, pft1-2hsr8-1 and pft1-2. Seedlings were grown vertically in the dark for 14 days on MS medium in the presence of 1 % Glucose. Errors bars represent SD from three biological replicates. Panels A-D **, p < 0.01 comparing Col to hsr8-1 (Student’s t- test); panels B and D **, p < 0.01 comparing pft1-2 hsr8-1 to pft1-2 (Student’s t- test). Relative transcript levels (RTL) were calculated using transcript levels of the reference gene TUB6 (At5g12250). (PPTX 97 kb)

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