Aspecto general del apéndice granulado al aire

Proyectos del Plan Nacional I+D+I con referencias PB94-0129, PB97-1132, BHA 2002-00138, HUM 2006-06250/HISTProyectos de la CAM con referencias 06/0020/1997, 06/0094/1998, 06/0090/2000, 06/0043/2001Programa Consolider-Ingenio 2010 con sigla CSD2007-00058NOMuseo Arqueológico Nacional (Madrid)GaleraArracada de racim

Additional file 16: Figure S2. of The complete chloroplast genome of Primulina and two novel strategies for development of high polymorphic loci for population genetic and phylogenetic studies

The PIC values of eight newly developed chloroplast markers calculated by bioinformatic analysis and experimental evaluation (PDF 222 kb

Kindlin-3 regulates the integrin αMβ2-Syk-Vav1 signaling axis.

<p>KM, ctrl-KM and k3-KM cells were seeded into iC3b-coated TC dishes in the presence of irrelevant mouse IgG (IgG) or mAb KIM185 (10 µg/ml each) and incubated under culture conditions for 30 min. Cells were harvested and lysed followed by immunoprecipitation (IP) using either anti-Syk or anti-Vav1 antibody with mouse IgG or rabbit anti-GST antibody as irrelevant IgG, respectively. Tyr-phosphorylated Syk and Vav1 were probed as described under <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0056911#s2" target="_blank">materials and methods</a>. IB: immunoblotting. A representative experiment from two independent experiments is shown.</p

Knockdown of kindlin-3 expression in K562 cells expressing integrin αMβ2.

<p>(A) qPCR analyses of kindlin-3 mRNA expression level in ctrl-KM and k3-KM cells. (B) Expression levels of kindlin-3 and other proteins in these cells were determined by immunoblotting. Actin was used as loading control. (C) Cell surface expression of integrin αMβ2 was determined by flow cytometry. Shaded and open histograms represent control IgG and mAb LPM19c stainings, respectively. GP: gated positive; GM: geo-mean; EI: expression index. (D) To determine extracellular activation of integrin αMβ2 on ctrl-KM and k3-KM cells. Cells were treated with Mn²⁺ (1 mM) or without and stained with mAb KIM127 at 37°C. Control IgG (ctrl-IgG) and mAb LPM19c were included for each condition. The %GP, GM and EI of mAb KIM127 staining are shown. One representative experiment out of two independent experiments is shown.</p

Kindlin-3 is required for integrin αMβ2-mediated cell spreading.

<p>ECIS measurements of ctrl-KM and k3-KM cells spreading on iC3b or BSA. Each data point represents the mean ± SD of technical triplicates at 1 min intervals. mAbs LPM19c and KIM185 were used at 10 µg/ml each. A plot of a representative experiment from three independent experiments is shown for each ligand.</p

Reduced kindlin-3 expression diminished integrin αMβ2-mediated cell adhesion.

<p>(A) and (B) show adhesion data of ctrl-KM and k3-KM cells on iC3b and BSA, respectively. Each data point represents the mean ± SD of three independent experiments. mAbs LPM19c and KIM185 were used at 10 µg/ml each. (C) Shear flow analyses of ctrl-KM and k3-KM cells in flow chambers coated with iC3b. Each data point is the mean ± SD of number of cells in four fields and a representative plot of two independent experiments is shown.</p

Kindlin-3 is involved in integrin αMβ2 outside-in signaling.

<p>(A) Flow cytometry analyses of αMβ2N329S in cells transduced with control or kindlin-3-targeting siRNA. Shaded and open histograms represent control IgG and LPM19c stainings, respectively. GP: gated positive; GM: geo-mean; EI: expression index. (B) Cell adhesion assay on iC3b. (C) ECIS measurements on iC3b. In (B) and (C), each data point represents mean ± SD of technical triplicates. mAb LPM19c was used at 10 µg/ml. (A–C), a single representative experiment from three independent experiments is shown.</p

Integrin αMβ2-induced RhoGTPase activation involves kindlin-3.

<p>Ctrl-KM and k3-KM cells were allowed to adhere to iC3b-coated TC dishes in the presence of mAb KIM185 (10 µg/ml). (A), (C) and (E) are immunoblots of cell lysates for Rac1, Cdc42 and RhoA, respectively. (B), (D) and (F) are pull-down experiments using cell lysates and RBD or PBD-conjugated beads. IB: immunoblotting. A representative experiment from two independent experiments is shown.</p

DataSheet1_Synergistic effect of CD47 blockade in combination with cordycepin treatment against cancer.docx

Cordycepin is widely considered a direct tumor-suppressive agent. However, few studies have investigated as the effect of cordycepin therapy on the tumor microenvironment (TME). In our present study, we demonstrated that cordycepin could weaken the function of M1-like macrophages in the TME and also contribute to macrophage polarization toward the M2 phenotype. Herein, we established a combined therapeutic strategy combining cordycepin and an anti-CD47 antibody. By using single-cell RNA sequencing (scRNA-seq), we showed that the combination treatment could significantly enhance the effect of cordycepin, which would reactivate macrophages and reverse macrophage polarization. In addition, the combination treatment could regulate the proportion of CD8+ T cells to prolong the progression-free survival (PFS) of patients with digestive tract malignancies. Finally, flow cytometry validated the changes in the proportions of tumor-associated macrophages (TAMs) and tumor-infiltrating lymphocytes (TILs). Collectively, our findings suggested that the combination treatment of cordycepin and the anti-CD47 antibody could significantly enhance tumor suppression, increase the proportion of M1 macrophages, and decrease the proportion of M2 macrophages. In addition, the PFS in patients with digestive tract malignancies would be prolonged by regulating CD8+ T cells.</p

Expression of kindlin-3 in melanoma cells impedes cell migration and metastasis

<p>Kindlins are a small family of 4.1-ezrin-radixin-moesin (FERM)-containing cytoplasmic proteins. Kindlin-3 is expressed in platelets, hematopoietic cells, and endothelial cells. Kindlin-3 promotes integrin activation, clustering and outside-in signaling. Aberrant expression of kindlin-3 was reported in melanoma and breast cancer. Intriguingly, kindlin-3 has been reported to either positively or negatively regulate cancer cell metastasis. In this study, we sought to clarify the expression of kindlin-3 in melanoma cells and its role in melanoma metastasis. Two widely used metastatic mouse and human melanoma cell lines B16-F10 and M10, respectively, were examined and found to lack kindlin-3 mRNA and protein expression. When kindlin-3 was ectopically expressed in these cells, cell migration was markedly reduced. These are attributed to aberrant Rac1 and RhoA activation and overt membrane ruffling. Our data demonstrate for the first time that despite its well established role as a positive regulator of integrin-mediated cell adhesion, aberrant expression of kindlin-3 could lead to imbalanced RhoGTPases signaling that impedes rather than promotes cell migration.</p