Additional file 1 of The synergistic mechanism of fibroblast growth factor 18 and integrin β1 in rat abdominal aortic aneurysm repair

Additional file 1: Fig. S1. Construction of the AAA model. (a) The abdominal aorta between the renal and iliac arteries was exposed and isolated from surrounding tissues using sterile rubber strips. (b, c) The abdominal aorta was wrapped in sterile cotton balls soaked in CaCl2 solution (0.8 mol/L). (d) Aneurysmal dilatation of the abdominal aorta. Figure S2. (a) Lentiviral transfection of ECs and SMCs: the fluorescence abundance of the LV-Con-RNAi group was higher than that of the LV-Itgβ1-RNAi group, whereas the fluorescence abundance of the LV-Itgβ1 group was higher than that of the LV-Con group. The fluorescence abundance of the NC + LV-Itgβ1 group was the highest, whereas that of the NC + LV-Itgβ1-RNAi group was the lowest. (b, c) WB and RT-qPCR assay for detecting Itgβ1 expression in ECs: Itgβ1 expression was lower in the NC + LV-Itgβ1-RNAi group than in the NC group; Itgβ1 expression was higher in the NC + LV-Itgβ1 group than in the NC group; Itgβ1 expression in the NC group was similar to that in the NC + LV-Con-RNAi and NC + LV-Con groups. (d, e) WB and RT-qPCR assay for detecting Itgβ1 expression in SMCs: Itgβ1 expression was lower in the NC + LV-Itgβ1-RNAi group than in the NC group; the Itgβ1 expression in the NC + LV-Itgβ1 group was higher than that in the NC group. **p < 0.01 versus NC group, ***p < 0.05 versus NC group. Figure S3 Transwell chamber assay for detecting the migratory activity of ECs and SMCs. (a) Both FGF18 and Itgβ1 could enhance the migratory activity of ECs; there was no significant difference in the migratory activity of ECs between the NC and NC + LV-Con groups. Moreover, the migratory activity of ECs in the NC + LV-Itgβ1-RNAi group was the weakest, whereas it was strongest in the NC + LV-Itgβ1 + FGF18 group. (b) The migratory activity of SMCs was higher in the NC + LV-Itgβ1 group than in the NC and NC + LV-Con groups. There was no significant difference in the migratory activity of SMCs between the NC and NC + LV-Con groups. Moreover, the migratory ability of SMCs was higher in the NC + LV-Itgβ1 + FGF18 group than in the other groups; bar = 200 μm

Additional file 1: Figure S1. of Retinoic acid promotes expression of germline-specific genes in chicken blastoderm cells by stimulating Smad1/5 phosphorylation in a feeder-free culture system

The mRNA expression levels of Smad1 and Smad5 were measured after RA, Dorso and SB431542 treatment. Data are representative of results in three independent experiments, values are the mean ± SEM (n = 3) and each condition is normalized to β-actin. *, p < 0.05; **, p < 0.01. (TIF 8187 kb

Crystal Structure of an Invasivity-Associated Domain of SdrE in <i>S</i>. <i>aureus</i>

<div><p>The surface protein SdrE, a microbial surface components recognizing adhesive matrix molecule (MSCRAMM) family protein expressed on the surface of <i>Staphylococcus aureus</i> (<i>S</i>. <i>aureus</i>), can recognize human complement regulator Factor H and C4BP, thus making it a potentially promising vaccine candidate. In this study, SdrE^278-591 was found to directly affect <i>S</i>. <i>aureus</i> host cell invasion. Additionally, the crystal structure of SdrE^278-591 at a resolution of 1.25 Å was established, with the three-dimensional structure revealing N2-N3 domains which fold in a manner similar to an IgG fold. Furthermore, a putative ligand binding site located at a conserved charged groove formed by the interface between N2 and N3 domains was identified, with β2 suspected to occupy the ligand recognizing site and undergo a structural rearrangement to allow ligand binding. Overall, these findings have further contributed to the understanding of SdrE as a key factor for <i>S</i>. <i>aureus</i> invasivity and will enable a better understanding of bacterial infection processes.</p></div

<i>S</i>. <i>aureus</i> SdrE^278-591 (magenta) superimposed on its homolog <i>S</i>. <i>aureus</i> Bbp (cyan).

<p>(A) <i>S</i>. <i>aureus</i> SdrE^278-591 (magenta) superimposed on its homolog Bbp (substrate free, PDB code 5cf3). (B) <i>S</i>. <i>aureus</i> SdrE^278-591 (magenta) superimposed on its homolog Bbp (substrate, PDB code 5cfa). The predominantly green strand is the peptide ligand (substrate).</p

Adherence and invasion of the ΔSdrE and ΔSdrE^278-591 mutants in host cell lines <i>in vitro</i>.

<p><i>S</i>. <i>aureus</i> Mu50 and its isogenic mutants <i>ΔSdrE</i> (Mu50Δ<i>SdrE</i>) and ΔSdrE^278-591 (Mu50Δ<i>SdrE-A</i>) were examined for adherence in HeLa (A) and 143B cells (B). These same mutants were also examined for invasivity in HeLa (C) and 143B (D) cells. Infectivity assessments were conducted for 4 h at 37°C. Wild-type <i>S</i>. <i>aureus</i> and the <i>ΔSdrE</i> (Mu50<i>ΔSdrE</i>) and <i>ΔSdrE</i>^278-591(Mu50<i>ΔSdrE-A</i>) mutants were generated to be devoid of SdrE in order to avoid destruction of the monolayer infection system. Scoring of the number of adherent and invasive bacterial cells indicate that adhesion and invasion are substantially reduced for <i>ΔSdrE</i> ((Mu50<i>ΔSdrE</i>) and <i>ΔSdrE</i>^278-591 -deficient (Mu50<i>ΔSdrE-A</i>) <i>S</i>. <i>aureus</i> mutant. Results are presented as a mean ± standard deviation for at least three independent experiments. Asterisks and triangles denote values significantly different from the wild-type as determined by Student’s t-test (** P < 0.01).</p

Data collection and refinement statistics.

<p>Data collection and refinement statistics.</p