Additional file 2: Table S2. of Transcriptome profiling of whitefly guts in response to Tomato yellow leaf curl virus infection

Sequences of primers used in the study. Displayed are the sequence of the primers used for qRT-PCR. Primers were synthesized by GenScript (Nanjing, China). (XLSX 9 kb

S100A4 expression in human TAA tissue and blood.

<p>A, Representative images of morphological and immunohistochemical analysis of a human TAA sample. The slides were stained with Weigert solution (a) (f), hematoxylin/eosin (b) (g), anti-S100A4 antibody (c) (h), anti-MMP-2 antibody (d) (i) and anti-MMP-9 antibody (e) (j). Scale bar = 100 µm. B, Higher power micrographs of the localization of S100A4 in the aortic intima (k), media (l) and adventitia (m). Scale bar = 50 µm. C, Concentrations of S100A4 determined by ELISA in serum obtained from TAA patients (n = 6) and control patients (n = 6). Values are the means ± SEM. *<i>P</i><0.05, indicates a significant difference from control patients.</p

Increased aortic diameter at all time-points after thoracic aortic aneurysm induction.

<p>A, Representative lower power micrographs of hematoxylin/eosin-stained, NaCl-treated aorta (control group) and CaCl₂-treated aorta (TAA group). Scale bar = 500 µm. B, Measurements of external media diameter in the control group and TAA group at all time-points. Bars represent the SEM, *<i>P</i><0.05 compared with the control group at each time-point (n = 6 for each group).</p

Down-regulation of S100A4 inhibited cell proliferation of A7r5 and MMP2/MMP9 expression <i>in vitro</i>.

<p>A, The results of cell proliferation ability of A7r5 cells after transfection with S100A4 siRNA at different time intervals (0, 24, 48, 72 h). *Represents <i>P</i><0.05, compared to the control nonspecific siRNA-transfected group and the untreated A7r5 group at 24, 48, and 72 h. B, RT-PCR analysis of MMP2 mRNA isolated from A7r5 cells from 3 groups after transfection at different time intervals (0, 24, 48, 72 h). * and # represents <i>P</i><0.05, compared to the control nonspecific siRNA-transfected group and the untreated A7r5 group at 24, 48, and 72 h. C, RT-PCR analysis of MMP9 mRNA isolated from A7r5 cells from 3 groups after transfection at different time intervals (0, 24, 48, 72 h). * and # represents <i>P</i><0.05, compared to the control nonspecific siRNA-transfected group and the untreated A7r5 group at 24, 48, and 72 h. D, Western blot analysis of the knockdown efficiency of S100A4 by siRNA. E, Western blot analysis of MMP2/MMP9 protein isolated from A7r5 cells from 3 groups after transfection at different time intervals (0, 24, 48, 72 h).</p

Representative pictures of immunohistochemical staining in aortic slides over time post-TAA induction.

<p>The slides were stained with antibodies against MMP-2, MMP-9, S100A4 and αSMA. An anti rabbit HRP/DAB detection system was used to visualize expression (brown staining). Scale bar = 50 µm.</p

Increased transcription levels of MMP-2, MMP-9 and S100A4A at all time-points after thoracic aortic aneurysm induction.

<p>A, Real-time PCR examination of MMP-2, MMP-9 and S100A4 at each time-point in the two groups, and mRNA levels are normalized to the mRNA of <i>β</i>-actin. Bars represent the SEM, *<i>P</i><0.05 compared with control group (n = 6 for each group). B, Positive correlations between the mRNA levels of S100A4 and MMPs over time post-TAA induction.</p

Disruption of the elastic lamellar structure at all time-points after thoracic aortic aneurysm induction.

<p>A, Representative Weigert staining and hematoxylin/eosin staining of sections of the aorta from NaCl-treated and TAA-induced rats. Scale bar = 100 µm. B, Grading of elastin degradation at each time-point in the two groups. C, Quantitation of intima-media thickness (IMT) in the two groups. Bars represent the SEM, *<i>P</i><0.05 compared with the control group (n = 6 for each group).</p

Spatiotemporal Expression of Matrix Metalloproteinases (MMPs) is Regulated by the Ca²⁺-Signal Transducer S100A4 in the Pathogenesis of Thoracic Aortic Aneurysm

<div><p>Aims</p><p>This study investigated whether S100A4 plays a potential role in the formation of thoracic aortic aneurysm (TAA).</p><p>Methods and Results</p><p>The thoracic aortas of male Sprague-Dawley rats were exposed to 0.5 M CaCl2 or normal saline (NaCl). Animals were euthanized at specified time-points (2, 4, and 10 weeks post-TAA induction). The treated aortic segments were harvested, and mRNA levels, protein expressions and immunohistochemistry of MMP-2, MMP-9 and S100A4 were analyzed. The A7r5 cell lines were used for an in vitro study. Experiments were also performed using human TAA samples for comparison. Localized aneurysmal dilation was observed in the CaCl2-treated segments. The transcription levels of S100A4 and MMPs were elevated in CaCl2-treated segments versus controls, and a significant correlation between S100A4 and expression of MMPs was observed across all time-points. Immunohistochemical studies revealed similar expression pattern of S100A4 and MMP proteins, as well as co-localization of S100A4 with the cell lineage markers (αSMA and CD68) and inflammatory markers (MMPs and NF-κB P65 subunit). The proliferative ability of A7r5 cells after transfection with S100A4 siRNA was suppressed, and down-regulation of S100A4 inhibited MMP-2 and MMP-9 expression in vitro. Increased expression of S100A4 was observed in all layers of the aorta wall in human TAA specimens. Serum concentrations of S100A4 determined by ELISA were found to be significantly increased in TAA patients.</p><p>Conclusions</p><p>This study established the important roles of S100A4 and MMPs in the development of TAA.</p></div

Clinical Characteristics of TAA Patients and Control Patients.

<p>Clinical Characteristics of TAA Patients and Control Patients.</p

Quantitation of the protein content of MMP-2, MMP-9, S100A4 and αSMA in aortic slides over time post-TAA induction.

<p>The immunohistochemically stained slides were assessed by computerized planimetry in the aortic adventitia (A, B and C) and aortic media (D, E, F and G). Bars represent the SEM, *<i>P</i><0.05 compared with control group (n = 6 for each group).</p