Syntax score according to HbA1c levels in type 2 DM patients aged 60 years and older.

<p>Syntax score according to HbA1c levels in type 2 DM patients aged 60 years and older.</p

Kaplan-Meier event-time curve according to preoperative NT-pro-BNP levels.

<p>Kaplan-Meier event-time curve according to preoperative NT-pro-BNP levels.</p

Risk factors associated with MACE in a multivariable logistic regression analysis.

<p>COPD: chronic obstructive pulmonary disease; ACE: angiotensin converting enzyme inhibitor; ARB: angiotensin receptor blocker.</p><p>Risk factors associated with MACE in a multivariable logistic regression analysis.</p

Baseline characteristics and laboratory data of the type 2 DM patients aged 60 years and older.

<p>LDL: low density lipoprotein; HDL: high density lipoprotein.</p

Kaplan-Meier survival curves according to preoperative NT-pro-BNP and cTnI levels

<p>Kaplan-Meier survival curves according to preoperative NT-pro-BNP and cTnI levels</p

Correlation between HbA1c levels and Syntax score in type 2 DM patients aged 60 years and older.

<p>Correlation between HbA1c levels and Syntax score in type 2 DM patients aged 60 years and older.</p

Opposite regulation of downstream genes in BR signaling by LIC and BZR1.

<p>(A) Transcriptional expression patterns of <i>ILI1</i> and <i>IBH1</i> in the <i>LIC</i> antisense line (AS2). Data are mean ± SD (<i>n</i> = 3). (B) Gel shift assay to illustrate LIC binding to the different fragments of the <i>ILI1</i> and <i>IBH1</i> promoters. <i>ILI1</i> B2 and <i>IBH1</i> C3 contain the binding element S. <i>ILI1</i> B1, B3, <i>IBH1</i> C1 or C2 fragments contain no or less binding elements. (C) and (D) ChIP analysis of LIC binding to the <i>ILI1</i> and <i>IBH1</i> promoters by use of anti-LIC antibody. The binding was enhanced in the <i>lic-1</i> mutant and in wild-type plants in the presence of BR. The black circle with white ring indicates the binding element S. B1–3 and C1–3 are the probes used in (B), and a–n (used in (C)) and a–k (used in (D)) indicate the sequences tested in ChIP assay. The <i>UBQUITIN5</i> promoter was used as a control. (E) Transient transfection assay to illustrate that LIC repressed <i>ILI1pro:LUC</i> and activated <i>IBH1pro:LUC</i> reporter gene expression in <i>Arabidopsis</i> protoplasts (the 403-bp <i>ILI1</i> promoter indicated as B2 in (C) and the 451-bp <i>IBH1</i> promoter indicated as C3 in (D) were used). The inhibition of <i>AtCPDpro:LUC</i> reporter gene expression by BZR1 was the control. Data are mean± SD. (F) Transient transfection assay indicated that LIC and BZR1 antagonistically regulate <i>ILI1pro:LUC</i> reporter gene expression. Data are mean ± SD.</p

LIC accumulates in the nucleus in response to BR treatment.

<p>(A) LIC accumulated in the nucleus in response to BR induction: (a) and (b) LIC-GFP fusion protein localized in both the nucleus and cytoplasm; LIC-GFP fluorescence intensity was weakened in the cytoplasm and enhanced in the nucleus with 1 µM 24-eBL treatment for 30 min. (c) and (d) LICm (mimic of <i>lic-1</i>) accumulated in the nucleus after 1 µM 24-eBL treatment similar to the intact LIC protein pattern. Numbers in each image show the mean signal of the total cell (1000×) and standard errors calculated from 10 cells for each treatment. The white lines inside the images show the areas used for line scan measurements that yielded plot profiles shown in the lower panels. The table shows signal intensities (10⁵×) and the ratios between nuclear and cytoplasmic (N/C) from represented areas. N, nuclear signal; C, cytoplasmic signal. The scale bar is 20 µm. (B) LICp-GFP fusion protein (deletion of both the C-terminus and P site) localized in only the nucleus. Bars = 20 µm. (C) A diagram for LIC protein (containing CCCH domain, EELR, P site and C terminus), LICm (deletion of the C-terminus, mimic of <i>lic-1</i>) and LICp (deletion of both the C-terminus and phosphorylation sites). (D) Immunoblotting analysis of LIC and LICm protein levels in the nuclear fractions and total protein. LICm localization was more in the nucleus, which is similar to wild-type LIC in BR-treated (1 µM) plants. LIC levels in the total protein did not change under the same condition. Histone 3 was the loading control for the nuclear fraction and Rubisco small subunit was the loading control for total protein.</p

LIC and BZR1 expression patterns and their responses to BR.

<p>(A) RNA <i>in situ</i> expression of <i>LIC</i> and <i>BZR1</i> on the abaxial and adaxial sides of leaves (the bottom panel represents the negative control with sense probes). Bar = 10 µm. (B) <i>LIC</i> and <i>BZR1</i> transcriptional expression response to various concentrations of BR. Data are mean ± SD (<i>n</i> = 5). *P<0.05 and **P<0.01 compared with no BR treatment as determined by Student's <i>t</i> test. (C) Immunoblotting to show the response of LIC protein expression to BR. LIC was repressed by low concentrations of BR (<100 nM) and induced by high concentrations of BR (>200 nM). Coomassie Blue staining served as the loading control. The levels of LIC were calculated after normalization against the intensity of Coomassie Blue staining in 3 replicated experiments, and the quantified values are shown beneath the gel images. Data are mean ± SE. (D) <i>LIC</i> transcriptional expression with BR treatment in wild-type (WT) and BR-deficient mutant <i>d2</i> and <i>BZR1</i> RNAi transgenic lines (<i>BZR1</i>R). <i>LIC</i> antisense line 2 (AS2) was a control. Data are mean ± SD (<i>n</i> = 3). (E) Time course response of transcription expression of <i>LIC</i> to BR (1 µM). <i>LIC</i> was rapidly induced by BR. Data are mean ± SD (<i>n</i> = 3). (F) <i>BZR1</i> and <i>CPD</i> transcriptional response to BR treatment in the wild type and <i>LIC a</i>ntisense lines. For <i>BZR1</i>, data are mean ± SD (<i>n</i> = 5). *P<0.05, compared with no BR treatment. For <i>CPD</i>, Data are mean ± SD (<i>n</i> = 3).</p

LIC is phosphorylated by BIN2/GSK1.

<p>(A) Immunoblotting analysis to demonstrate that LIC was phosphorylated by BIN2. LIC phosphorylation was antagonized by λ-phosphatase 1 (PP1). The phosphorylation status of LIC is illustrated by autoradiography of an anti-LIC antibody in the top panel. The amount of protein is shown with Coomassie Blue staining in the bottom panel. The levels of unphosphorylated LIC relative to the control without BIN2 and PP1 (-P%) were calculated after normalization against the intensity of Coomassie Blue staining, and these values are shown beneath the gel images. (B) Treatment with BR (1 µM) decreased the levels of phosphorylated LIC and increased that of unphosphorylated LIC. Rice plants were grown for 2 weeks and then soaked with 1 µM 24-eBL (+) or mock solution (−) for 3 h. LIC protein was analyzed by immunoblotting with an anti-LIC antibody (upper panel). The loading control with Coomassie Blue staining is shown in the bottom panel. (C) The mutated protein LICm caused decreased phosphorylation in the <i>lic-1</i> mutant. The 24-eBL concentration was 1 µM. (D) Immunoblotting assay for LIC protein in the nuclear and cytoplasmic fractions. Dephosphorylated LIC was dominant in the nucleus (N), and phosphorylated forms were dominant in the cytoplasm. Nuclear and cytoplasmic protein fractions were extracted from 2-week-old rice seedlings. Histone 3 was a marker for the nuclear protein and ß-actin for the cytoplasmic protein.</p