Journal: Scientific Reports

Article Title: PPP1 , a plant-specific regulator of transcription controls Arabidopsis development and PIN expression

doi: 10.1038/srep32196

Figure Lengend Snippet: ( a) Alignment of PIN promoter fragments. Nucleotides identical to the 16 bp motif characterized in the PIN2 promoter are in red. A 7-nucleotide motif found in the promoters of PIN1 , PIN2 , PIN3 , PIN7 and PIN8 is highlighted in yellow. Sequence logo displaying the position frequency matrix of aligned sequences of PIN1 , PIN2 , PIN3 , PIN6 , PIN7 and PIN8 . ( b) Plot of all 16384 possible 7-mers matched to the promoter regions of 33217 Arabidopsis genes (x-axis), and to the promoters of all 8 Arabidopsis PIN genes (y-axis). For each 7-mer, a p-value is calculated based on the hypergeometric distribution, with hits with a p-value < 0.05 shown in black. The minimum 7-nucleotide common sequence found in the PIN promoters is depicted in red and circled.

Article Snippet: Antibodies were diluted as follows: 1:500 for rabbit anti-PIN1 , anti-PIN2 and 1:300 for FITC-conjugated anti-rabbit secondary antibodies (Dianova).

Techniques: Sequencing

Journal: Scientific Reports

Article Title: PPP1 , a plant-specific regulator of transcription controls Arabidopsis development and PIN expression

doi: 10.1038/srep32196

Figure Lengend Snippet: ( a ) qPCR analysis of PPP1 expression in wild type (“WT”) seedlings versus ppp1-411 insertion mutant seedlings (left) and an ami-ppp1 silencer used for further analysis (right; see text). Graphs depict expression fold change in PPP1 expression in ppp1-411 and ami-ppp1 relative to WT and normalized to the expression of 2 reference genes ( EIF4a and TUB ). Error bars depict s.e.m. from 3 biological replicates. ( b ) qPCR analysis of PIN1 and PIN2 transcript levels in wild type (WT) seedlings versus ami-ppp1 seedlings. Graph depicts expression fold change in PIN gene expression in the ami-ppp1 line relative to WT and normalized to the expression of 2 reference genes ( EIF4a and TUB ). Error bars depict s.e.m. from 3 biological replicates. ( c,d ) Activity of PIN2p::GUS in a wild type (c) and in a ami-ppp1 (d) root meristem. ( e,f ) Whole mount immuno-labeling performed with wild type (top) and ami-ppp1 (bottom) seedlings at 4 DAG that were probed with anti-PIN2 (e) and anti-PIN1 (f). ( g ) qPCR analysis of transcript levels of genes involved in intra- and intercellular auxin transport in wild type (WT) and ami-ppp1 seedlings. Graph depicts expression fold change in gene expression in the ami-ppp1 line relative to WT. Error bars depict s.e.m from 3 biological replicates. Bars: c,d = 100 μm; e,f = 20 μm.

Article Snippet: Antibodies were diluted as follows: 1:500 for rabbit anti-PIN1 , anti-PIN2 and 1:300 for FITC-conjugated anti-rabbit secondary antibodies (Dianova).

Techniques: Expressing, Mutagenesis, Activity Assay, Immunolabeling

Journal: Scientific Reports

Article Title: PPP1 , a plant-specific regulator of transcription controls Arabidopsis development and PIN expression

doi: 10.1038/srep32196

Figure Lengend Snippet: ( a,b ) Comparison of wild type (a) and ami-ppp1 (b) plantlets grown on vertically orientated agar plates for 11 days. Red arrowheads indicate direction of gravity vector ( c ) Wild type (left) and ami-ppp1 (right) plants at 35 days. ( d ) Primary root length of wild type and ami-ppp1 seedlings at 8 DAG. Error bars represent standard deviations (n = 30 seedlings for each genotype; statistical analysis was performed using Student’s two-tailed t-test). ( e ) Orientation of primary root growth of wild type and ami-ppp1 seedlings at 8 DAG. A total of 114 (wild type) and 118 ( ami-ppp1 ) seedlings was analyzed in 3 biological repeats, and plotted as percentage of seedlings displaying <15°, <30°, <45° and >45° deviation from the vertical growth axes. Error bars indicate standard deviations. ( f,g ) Comparison of ppp1-411 (f) and ppp1-411 35S::PIN1 (g) primary root meristems at 9 DAG. White arrowheads depict onset of cell elongation. ( h ) Partial rescue of ppp1-411 root elongation by 35S::PIN1 . Root lengths of wild type (WT), 35S::PIN1 , ppp1-411 and ppp1-411/35S::PIN1 . The latter is significantly longer compared to ppp1-411 mutants (n ≥ 20 individuals analyzed for each genotype; statistical analysis was performed using Student’s two-tailed t-test; standard deviations are shown as error bars). Bars: a,b = 10 mm; c = 50 mm; f,g = 50 μm.

Article Snippet: Antibodies were diluted as follows: 1:500 for rabbit anti-PIN1 , anti-PIN2 and 1:300 for FITC-conjugated anti-rabbit secondary antibodies (Dianova).

Techniques: Plasmid Preparation, Two Tailed Test