Table_1_Genome-wide identification and expression profiles analysis of the authentic response regulator gene family in licorice.xlsx

IntroductionAs one of the traditional Chinese medicinal herbs that were most generally used, licorice attracts lots of interest due to its therapeutic potential. Authentic response regulators (ARRs) are key factors in cytokinin signal transduction and crucial for plant growth and stress response processes. Nevertheless, the characteristics and functions of the licorice ARR genes are still unknown.ResultsIn present study, a systematic genome-wide identification and expression analysis of the licorice ARR gene family were conducted and 51 ARR members were identified. Collinearity analysis revealed the significant roles of segmental duplications in the expansion of licorice ARR genes. The cis-acting elements associated with development, stress and phytohormone responses were identified, implying their pivotal roles in diverse regulatory processes. RNA-seq and qRT-PCR results suggested that A-type, but not B-type ARRs were induced by zeatin. Additionally, ARRs participated in diverse abiotic stresses and phytohormones responses. Yeast one-hybrid assay demonstrated that GuARR1, GuARR2, GuARR11, GuARR12, GuARR10-1, GuARR10-2 and GuARR14 were able to bind to the promoter of GuARR8-3, and GuARR1, GuARR12 bound to the GuARR8-1 promoter. GuARR1, GuARR2, GuARR11 and GuARR10-2 bound to the GuARR6-2 promoter as well as GuARR12 and GuARR10-2 bound to the GuARR6-1 promoter.DiscussionCollectively, these findings provide a basis for future ARR genes function investigations, shedding light on the potential medicinal properties and agricultural applications of licorice.</p

Table_4_Genome-wide identification and expression profiles analysis of the authentic response regulator gene family in licorice.xlsx

IntroductionAs one of the traditional Chinese medicinal herbs that were most generally used, licorice attracts lots of interest due to its therapeutic potential. Authentic response regulators (ARRs) are key factors in cytokinin signal transduction and crucial for plant growth and stress response processes. Nevertheless, the characteristics and functions of the licorice ARR genes are still unknown.ResultsIn present study, a systematic genome-wide identification and expression analysis of the licorice ARR gene family were conducted and 51 ARR members were identified. Collinearity analysis revealed the significant roles of segmental duplications in the expansion of licorice ARR genes. The cis-acting elements associated with development, stress and phytohormone responses were identified, implying their pivotal roles in diverse regulatory processes. RNA-seq and qRT-PCR results suggested that A-type, but not B-type ARRs were induced by zeatin. Additionally, ARRs participated in diverse abiotic stresses and phytohormones responses. Yeast one-hybrid assay demonstrated that GuARR1, GuARR2, GuARR11, GuARR12, GuARR10-1, GuARR10-2 and GuARR14 were able to bind to the promoter of GuARR8-3, and GuARR1, GuARR12 bound to the GuARR8-1 promoter. GuARR1, GuARR2, GuARR11 and GuARR10-2 bound to the GuARR6-2 promoter as well as GuARR12 and GuARR10-2 bound to the GuARR6-1 promoter.DiscussionCollectively, these findings provide a basis for future ARR genes function investigations, shedding light on the potential medicinal properties and agricultural applications of licorice.</p

Table_3_Genome-wide identification and expression profiles analysis of the authentic response regulator gene family in licorice.xlsx

IntroductionAs one of the traditional Chinese medicinal herbs that were most generally used, licorice attracts lots of interest due to its therapeutic potential. Authentic response regulators (ARRs) are key factors in cytokinin signal transduction and crucial for plant growth and stress response processes. Nevertheless, the characteristics and functions of the licorice ARR genes are still unknown.ResultsIn present study, a systematic genome-wide identification and expression analysis of the licorice ARR gene family were conducted and 51 ARR members were identified. Collinearity analysis revealed the significant roles of segmental duplications in the expansion of licorice ARR genes. The cis-acting elements associated with development, stress and phytohormone responses were identified, implying their pivotal roles in diverse regulatory processes. RNA-seq and qRT-PCR results suggested that A-type, but not B-type ARRs were induced by zeatin. Additionally, ARRs participated in diverse abiotic stresses and phytohormones responses. Yeast one-hybrid assay demonstrated that GuARR1, GuARR2, GuARR11, GuARR12, GuARR10-1, GuARR10-2 and GuARR14 were able to bind to the promoter of GuARR8-3, and GuARR1, GuARR12 bound to the GuARR8-1 promoter. GuARR1, GuARR2, GuARR11 and GuARR10-2 bound to the GuARR6-2 promoter as well as GuARR12 and GuARR10-2 bound to the GuARR6-1 promoter.DiscussionCollectively, these findings provide a basis for future ARR genes function investigations, shedding light on the potential medicinal properties and agricultural applications of licorice.</p

Table_5_Genome-wide identification and expression profiles analysis of the authentic response regulator gene family in licorice.xlsx

IntroductionAs one of the traditional Chinese medicinal herbs that were most generally used, licorice attracts lots of interest due to its therapeutic potential. Authentic response regulators (ARRs) are key factors in cytokinin signal transduction and crucial for plant growth and stress response processes. Nevertheless, the characteristics and functions of the licorice ARR genes are still unknown.ResultsIn present study, a systematic genome-wide identification and expression analysis of the licorice ARR gene family were conducted and 51 ARR members were identified. Collinearity analysis revealed the significant roles of segmental duplications in the expansion of licorice ARR genes. The cis-acting elements associated with development, stress and phytohormone responses were identified, implying their pivotal roles in diverse regulatory processes. RNA-seq and qRT-PCR results suggested that A-type, but not B-type ARRs were induced by zeatin. Additionally, ARRs participated in diverse abiotic stresses and phytohormones responses. Yeast one-hybrid assay demonstrated that GuARR1, GuARR2, GuARR11, GuARR12, GuARR10-1, GuARR10-2 and GuARR14 were able to bind to the promoter of GuARR8-3, and GuARR1, GuARR12 bound to the GuARR8-1 promoter. GuARR1, GuARR2, GuARR11 and GuARR10-2 bound to the GuARR6-2 promoter as well as GuARR12 and GuARR10-2 bound to the GuARR6-1 promoter.DiscussionCollectively, these findings provide a basis for future ARR genes function investigations, shedding light on the potential medicinal properties and agricultural applications of licorice.</p

Table_2_Genome-wide identification and expression profiles analysis of the authentic response regulator gene family in licorice.xlsx

IntroductionAs one of the traditional Chinese medicinal herbs that were most generally used, licorice attracts lots of interest due to its therapeutic potential. Authentic response regulators (ARRs) are key factors in cytokinin signal transduction and crucial for plant growth and stress response processes. Nevertheless, the characteristics and functions of the licorice ARR genes are still unknown.ResultsIn present study, a systematic genome-wide identification and expression analysis of the licorice ARR gene family were conducted and 51 ARR members were identified. Collinearity analysis revealed the significant roles of segmental duplications in the expansion of licorice ARR genes. The cis-acting elements associated with development, stress and phytohormone responses were identified, implying their pivotal roles in diverse regulatory processes. RNA-seq and qRT-PCR results suggested that A-type, but not B-type ARRs were induced by zeatin. Additionally, ARRs participated in diverse abiotic stresses and phytohormones responses. Yeast one-hybrid assay demonstrated that GuARR1, GuARR2, GuARR11, GuARR12, GuARR10-1, GuARR10-2 and GuARR14 were able to bind to the promoter of GuARR8-3, and GuARR1, GuARR12 bound to the GuARR8-1 promoter. GuARR1, GuARR2, GuARR11 and GuARR10-2 bound to the GuARR6-2 promoter as well as GuARR12 and GuARR10-2 bound to the GuARR6-1 promoter.DiscussionCollectively, these findings provide a basis for future ARR genes function investigations, shedding light on the potential medicinal properties and agricultural applications of licorice.</p

Absolute counts of infiltrating DCs recruited to the burn and sham injury site.

<p>Three days post burn injury, the skin specimens of burn and sham injury (2 cm×2 cm) were harvested and digested in Dispase II and collagenase D. Cells were isolated and stained for CD11c⁺ for DCs followed by flow cytometry analysis. Absolute numbers of infiltrating CD11c⁺ DCs are shown as mean ± SEM (n<i> = </i>8, 4 independent experiments).</p

Reduced TLR9-mediated DC maturation following burn injury.

<p>Total splenic CD11c⁺ DCs were purified at different time point post-burn/sham injury and subjected to CpG activation. TLR-activated cells were stained for CD11c, B220, PDCA1, CD80, CD86 and MHC II expression and CDCs were gated as CD11c^hiB220⁻ and pDCs as CD11c^lowB220⁺PDCA1⁺. Percentages of MHCII^hiCD80^hiCD86^hi mature cDCs (a) and pDCs (b), as well as MFI of MHC-II expression on cDCs (a) and pDCs (b) are shown as mean ± SEM (n >9, 3–5 independent experiments). *<i>P</i><0.05, **<i>p</i><0.01, sham versus burn by ANOVA.</p

Reduced numbers and percentages of splenic cDCs and pDCs following burn injury.

<p>Mice were subjected to non-lethal thermal injury and total splenocytes were purified and stained using antibodies for distinct surface markers. The effect of burn injury on each DC subset was examined. (a) A representative FACS plot demonstrating the percentages of splenic cDCs (CD11c^hiB220^neg) and CD11c^lowB220⁺ DC subpopulations at d3 post-injury in comparison to sham. The CD11c^lowB220⁺ population comprises of pDCs (CD11c^lowB220⁺PDCA1⁺) and IKDCs (CD11c^lowB220⁺DX5⁺). Percentages of CD11c^hiB220^neg cDCs and CD11c^lowB220⁺ DCs are shown as mean ± SEM (n = 9, 3 independent experiments). Percentages (b) and absolute numbers (c) of splenic cDCs, pDCs and IKDCs at various time points post burn and sham injury are shown. Data are shown as mean ± SEM (n = 9, 3 independent experiments). *<i>P</i><0.05; **<i>p</i><0.01, sham versus burn by ANOVA.</p

CDCs and pDCs from burn-injured mice expressed a lower transcript level of TLR9 and genes related to TLR signaling pathway.

<p>Three days post-injury, cDCs and pDCs were purified by FACS sorting. (a) Transcript level of TLR9 was examined by real-time PCR. Fold changes of TLR expressions of burn and sham mice were normalized to those of untreated control mice. The data analyzed represents the mean ± SEM of 5–6 independent experiments using 10 mice per group. *, <i>p</i><0.05, sham versus burn by ANOVA. (b) The expressions of genes related to TLR-mediated signal transduction were examined by real-time PCR Array. Fold changes of transcript levels of genes in cDCs and pDCs of burn mice were normalized to the corresponding genes of sham mice per experiment. Genes expression in cDC and pDC of sham mice were set as 1. The data analyzed represent the mean ± SEM (n = 10, 2 independent experiments).</p

PDCs from burn-injured mice had an impaired ability to secrete pro-inflammatory cytokines.

<p>Three days post-injury, FACS-sorted spleen pDCs with purity >98% were activated with CpG (18–20 hr). Inflammatory cytokines and IFN-α productions by TLR9-activated pDCs were measured by CBA assay and ELISA, respectively. Data are shown as mean ± SEM (n = 8, 4 independent experiments). *<i>P</i><0.05, sham versus burn by ANOVA.</p