Fmn1-IV −/− MEFs exhibit reduced focal adhesion formation in a cell spreading assay.

<p>(A) Wild-type and Fmn1-IV null MEFs were replated on fibronectin substrates for 10 or 20 min before fixation. Focal adhesions were imaged by immunofluorescence after staining with phospho-tyrosine or phospho-caveolin antibodies. (B) The graph represents the percentages of cells (out of 100%) possessing the range of focal adhesions, as indicated. Numbers of cells examined for focal adhesions: Fmn1-IV +/+ 10 min n = 192, Fmn1-IV −/− 10 min n = 162, Fmn1-IV +/+ 20 min n = 176, Fmn1-IV −/− 20 min n = 167.</p

Intracellular localization of Fmn1-IV in primary kidney epithelial cells derived from EGFP-Fmn1-IV knock-in mice (A-L).

<p>Immunofluorescence staining of primary epithelial cells with anti-GFP antibody (A), anti-E-cadherin antibody (B), and phalloidin (C). Merged images: (D) anti-GFP and phalloidin (magnified insert, E), (F) anti-GFP and E-cadherin (magnified insert, G), and (H) anti-GFP, phalloidin and E-cadherin (magnified insert, I). Confluent epithelial cells stained with anti-GFP (J), anti-β-catenin (K), and merged image (L). Note that Fmn1-IV is largely absent from adherens junctions, by its lack of co-localization with E-cadherin and β-catenin.</p

Rabbit polyclonal antibody raised against exon 6 of Fmn1-IV predominately reacts with actin.

<p>(A) MEFs derived from Fmn1-IV wild type and null mice were stained for immunofluorescence with anti-Fmn1-IV antibodies <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0002497#pone.0002497-Kobielak1" target="_blank">[29]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0002497#pone.0002497-ORourke1" target="_blank">[31]</a> and phalloidin, as indicated at the top of the panels. Note that the staining patterns are similar for all images. (B) Western blot of MEF extracts stained with anti-Fmn1 antibody <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0002497#pone.0002497-Chan2" target="_blank">[32]</a>, and actin as a loading control. (C) Western blot of MEF extracts and actin (last lane) stained with anti-Fmn1-IV antibody <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0002497#pone.0002497-Kobielak1" target="_blank">[29]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0002497#pone.0002497-ORourke1" target="_blank">[31]</a>.</p

Fmn1-IV −/− MEFs exhibit reduced cell spreading behavior.

<p>(A) Wild-type and Fmn1-IV null MEFs were fixed and stained for filamentous actin after 15 min of replating on fibronectin. The total cell area was measured with Openlab. (B) Results shown are the mean±SEM (error bars). **, P<0.001.</p

Localization of EGFP-Fmn1-IV by immuno-gold EM in MEFs derived from knock-in mice.

<p>(A and B) Tubulin and EGFP are labeled with 6 nm and 15 nm gold particles, respectively. Open head arrows indicate microtubules, and arrows indicate EGFP-Fmn1-IV association along microtubules.</p

Fmn1-IV −/− MEFs exhibit altered protrusive behavior.

<p>Time-lapse microscopic images were taken from the peripheries of representative wild-type and Fmn1-IV −/− MEFs grown on a fibronectin substrate after wound healing. Kymographs of the leading edge of cells were analyzed both for protrusive and retractive behavior. (A) Protrusive persistence (Δx), (B) protrusive distance (Δy), (C) protrusion rate (Δy/Δx), and (D) retraction rate (−Δy/Δx) were determined and the data represent the mean±SEM (error bars). *, P<0.05.</p

Generation of the EGFP-Fmn1-IV fusion allele.

<p>(A) Diagram of the strategy for generating an in-frame insertion of EGFP into the Fmn1-IV locus, which produces an EGFP-tagged Fmn1-IV driven from the endogenous promoter in mice. The star designates the start of translation. The black filled boxes flanking the PGK Neo cassette represent LoxP sites. (B) Representative PCR genotyping of tail DNA from mice possessing the EGFP-Fmn1-IV allele are shown. The letter F indicates the fusion allele, while + indicates wt allele.</p

Formation of adherens junctions in Fmn1-IV null primary kidney epithelial cells.

<p>Primary kidney epithelial cells derived from Fmn1-IV wild type and null mice were stained for immunofluorescence with anti-E-cadherin antibodies.</p

Regulating the Mesoporous Structure of Carbon Nanospheres by a Local Ablation Method for High-Performance PEMFC Catalysts

Pt-based catalysts are the most widely used catalysts in proton exchange membrane fuel cells (PEMFCs). However, the catalytic activity cannot be fully expressed by the pore structure of current commercial catalysts. In this work, the pore structure of available commercial carbon black (4–7 nm), which is beneficial for Pt catalytic activity, was successfully regulated by using perchloric acid (HClO4) as a pore-forming agent. The generated carbon nanospheres are denoted as NCB (new carbon black), with a pore volume in the 4–7 nm region (V4–7 nm) increasing from 0.107 cm3/gcarbon to 0.164 cm3/gcarbon. Pt-embedded catalysts on NCB (Pt/NCB) were synthesized by the impregnation method. The Pt/NCB catalyst exhibited oxygen reduction reaction (ORR) activity comparable to that of the original porous carbon-supported Pt catalyst under the suppression of ionomers. Moreover, the proton conduction resistance of the Pt/NCB catalyst layer decreased from 78 Ω·cm2 to 57 Ω·cm2, a decrease of 26.9%. The Pt/NCB catalyst layer also displayed excellent oxygen transport performance, and the oxygen gain voltage (OGV) was lower than that of the Pt catalyst with commercial porous carbon support. Overall, the local ablation method with HClO4 as a pore-forming agent is an effective way to fabricate accessible pores on easily available commercial carbon black, contributing to highly efficient catalytic activity

Additional file 1 of Association between lactate-to-albumin ratio and 28-days all-cause mortality in patients with sepsis-associated liver injury: a retrospective cohort study

Additional file 1: Supplementary Table 1. Specific value and percentage of missing variable