Xwnt5A-EGFP but not ROR2-mCherry clusters in membranes of DMZ cells.

<p>A) 150 pg of Xwnt5A-EGFP mRNA were co-injected with 125 pg membrane-anchored (growth-associated protein 43 (Gap43))-mCherry as a lineage tracer in the two dorsal blastomeres of eight-cell stage embryos. At stage 10.25, dorsal marginal zones were explanted and analyzed for subcellular localization of the Xwnt5A-EGFP protein. Prominent Xwnt5A-EGFP clusters localized at the membrane of DMZ cells at the onset (stage 10.5) and end (stage 12) of gastrulation. Shown are snapshots of <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0109428#pone.0109428.s003" target="_blank">movies S1</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0109428#pone.0109428.s004" target="_blank">S2</a>. B) 40 pg ROR2-mCherry were injected into the two dorsal blastomeres of eight-cell stage embryos. ROR2-mCherry is homogenously distributed at the membrane of DMZ explants throughout gastrulation. Shown are snapshots of <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0109428#pone.0109428.s005" target="_blank">movies S3</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0109428#pone.0109428.s006" target="_blank">S4</a>.</p

Long-range Wnt-5A inhibits the ATF2-Luc reporter.

<p>A) For studying long-range signaling, Xwnt5A-EGFP transfected HEK293 cells seeded on thinCerts TC chambers were transferred on cells transfected with the ATF2-Luc reporter and cultivated for additional 24 hours. To investigate paracrine signaling, cells transfected with Xwnt5A-EGFP were co-cultured with cells transfected with the ATF2-Luc reporter for 24 hours. To analyze both autocrine and paracrine signaling, the reporter and the Wnt ligand were cotransfected. B) Activation of the non-canonical Wnt reporter ATF2-Luc by co-transfected Wnt8, Wnt11 and Wnt5A. C) Activation of the non-canonical Wnt reporter ATF2-Luc by long-range Wnt8, Wnt11 and Wnt5A in a two-chamber assay. D) Xwnt5A-EGFP triggered ATF2-Luc activation depends on endocytosis. Wnt5A induced ATF2-Luc activation (cotransfection) was inhibited by addition of 5 µg/ml chlorpromazine (Chlo) and 150 µM genistein (Gen) 24 h before the measurement. Given are the mean values ± standard errors and the p-values according to Student's t-test, ns: not significant, n gives the number of transfections.</p

Xwnt5A induces clustering of ROR2.

<p>A) 150 pg of Xwnt5A-EGFP mRNA were co-injected with 40 pg of the ROR2-mCherry in the two dorsal blastomeres of eight-cell stage embryos. At stage 10.25 dorsal marginal zones were explanted and analyzed for subcellular localization of the Xwnt5A-EGFP and ROR2-mCherry. Prominent Xwnt5A-EGFP clusters co-localize with ROR2-mCherry clusters at the membrane of DMZ cells. These clusters are found at the onset (stage 10.5) and at the end (stage 12) of gastrulation. Shown are snapshots of <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0109428#pone.0109428.s007" target="_blank">movies S5</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0109428#pone.0109428.s008" target="_blank">S6</a>. B) 150 pg of Xwnt5A-EGFP mRNA were co-injected with 1.6 pmol Xwnt-5A specific antisense morpholino and 125 pg Gap43-mCherry. Also in these Xwnt-5A-depleted explants Xwnt5A-EGFP clustered in the membrane.</p

Supramolecular Self-Assembly Bioinspired Synthesis of Luminescent Gold Nanocluster-Embedded Peptide Nanofibers for Temperature Sensing and Cellular Imaging

Metal nanoclusters (NCs) hold great potential as novel luminescent nanomaterials in many applications, while the synthesis of highly luminescent metal NCs still remains challenging. In this work, we report self-assembling peptides as a novel bioinspired scaffold capable of significantly enhancing the luminescence efficiency of gold nanoclusters (AuNCs). The resulting AuNCs capped with motif-designed peptides can self-assemble to form nanofiber structures, in which the luminescence of AuNCs is enhanced nearly 70-fold, with 21.3% quantum yield. The underlying mechanism responsible for the luminescence enhancement has been thoroughly investigated by the combined use of different spectroscopic and microscopic techniques. The resultant highly luminescent AuNC-decorated peptide nanofibers exhibit physicochemical properties that are advantageous for biological applications. As a proof of concept, we demonstrate the use of these nanostructure as fluorescent thermometers and for imaging living cells, both showing very promising results

Data from Mattes 2018

Kinase Screen and automated image-analysis identifies the receptor tyrosine kinase Ror2 as a potential cytoneme regulator upstream of Cdc42. Wnt8a-GFP and a membrane bound mCherry was co-transfected with kinase library genes in Pac2 cells in a 96-well plate. Images were acquired automatically and analyzed for filopodia length and numbers by a filopodia detection software. Automated Image analysis software detects and counts filopodia of single cells using the memCherry signal and quantifies their length by automatically tracing the tips back to the cell body. Table shows transfected kinase genes and their relative filopodia number/cell and length