Research on the Influence of Convenience Store Brand Image on Brand Loyalty from the Perspective of Brand Identity

亜細亜大学修士（経営学）thesi

イマデガワ キャンパス セイビ ニ トモナウ リッカイ チョウサ セイカ(2009～2012ネンド)

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Observation of b→dγ and Determination of |Vtd/Vts|

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「弘長百首」について

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Process extension by solitary HB1.F3.eGFP cells does not vary with target cell type.

<p><b><i>A</i></b>, Illustration of Scholl analysis with concentric rings at 10 µm spacing centered and superimposed on the HB1.F3.eGFP cell soma. <b><i>B</i></b>, Proportions of HB1.F3.eGFP cells exhibiting processes after culture in the absence of target cells, and after co-culture with potential target cells: glioma (U251, U87, D556); breast cancer (MCF-7, SK-BR3, MDA-MB-231); fibroblast (3T3), primary mouse astroglial cells. No significant differences (all <i>p</i>>0.05) were seen between target cells. <b><i>C</i></b>, Lengths of HB1.F3.eGFP cell process after co-culture with each of the target cells indicated are virtually identical. Data are mean ± s.d. for 3 fields for 2–4 independent experiments.</p

Analyses of HB1.F3.eGFP::target cell contacts.

<p>Formation of contacts, and the extent of target cell encirclement, did not vary with types of cell encountered, including multiple human malignant lines, primary human glioma cell lines, primary mouse astroglia, and human dermal fibroblasts (adult and new born) and astrocytes. <b><i>A</i></b>, Proportions of total HB1.F3.eGFP cells contacting target cells. U251 cells were one of several malignant brain and breast cancer lines analyzed, and in addition served as a reference cell line for each additional group (marked by either dsRed or CMRA as indicated). No significant differences were observed (all <i>p</i>>0.05 within each group of target cells and across multiple groups; ANOVA with Bonferroni's multiple comparisons test, or <i>t</i> test, as appropriate). <b><i>B</i></b>, Proportions of HB1.F3::target cell pairs displaying each of the thee morphologies defined as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0051859#pone-0051859-g003" target="_blank">Figure 3</a> and illustrated in the legend (showing only the HB1.F3.eGFP cell). No significant differences were observed (all <i>p</i>>0.05) with the exception of HB1.F3 homotypic interactions (see text). Data are mean ± s.d. for 3 or more volumes within each of 2–4 independent experiments.</p

Stages of HB1.F3 contact formation and encirclement of U251.dsRed cells.

<p>Images are Z-axis projections of 28–44 optical sections taken at 1 µm intervals. <b><i>A</i></b>, Solitary HB1.F3.eGFP cell extending filopodia-tipped process into the hydrogel matrix. <b><i>B</i></b>, Initial HB1.F3.eGFP contact with a U251.dsRed cell (<b>Type I</b>). Note the morphology of the process contacting the target cell, which has shorter stubbier extensions resembling a holdfast. <b><i>C</i></b>, Loss of processes, apposition of the HB1.F3.eGFP cell body to the U251.dsRed cell, and the beginnings of lamellipodia extension (<b>Type II</b>). <b><i>D</i></b>, Complete HB1.F3.eGFP cell encirclement of a U251.dsRed cell with the cell body and lamellipodia covering much of the glioma cell surface (<b>Type III</b>).</p

Spatial relationships between HB1.F3.eGFP NSCs (green) and U251.dsRed glioma cells (red).

<p>Cells were co-cultured for 18 h in 3-dimensional Puramatrix hydrogel. Shown is the Z-axis projection of 34 optical sections of 900×900 µm spanning 102 µm. Expanded X-Z projections of individual contacting cell pairs confirm true contacts rather than superposition. Of a total of 157 HB1.F3.eGFP cells in this image, 38 (36.9%) were contacting U251.dsRed cells. Of the remaining solitary cells, 94 (59.9%) were roughly spherical or ellipsoid with minimal or no process extension, and 25 (15.9%) showed extended filopodia-tipped processes.</p

HB1.F3.eGFP cells will form contacts with, but not encircle, other HB1.F3 cells.

<p><i>Above</i>, Images of HB1.F3.eGFP::HB1.F3.eGFP and HB1.F3.eGFP::HB1.F3.CD (CMRA) contacting pairs. Cell boundaries are outlined using phalloidin_Alexa647 to show the cortical F-actin cytoskeleton; contact zones are marked by arrows. While the HB1.F3 cells showed filopodial extensions over their hydrogel-exposed surfaces, the contact zone was smooth and clearly delineated by the phalloidin_Alexa647. <i>Below</i>, Heterotypic HB1.F3.eGFP::U251.dsRed pairs were characterized by extension of actin-rich (arrows) lamellipodia from the HB1.F3.eGFP cell over the target glioma cell. These images illustrate the data presented quantitatively in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0051859#pone-0051859-g005" target="_blank">Figure 5</a>.</p

Disruption of F-actin filaments with cytochalasin D and myosin II by blebbistatin and consequences for motor protein distributions and HB1.F3.eGFP interaction with target U251.dsRed cells.

<p><b><i>A</i></b>, Images illustrating the sensitivities of cell morphology in Puramatrix (<i>a1, b1, c1</i>) and of the distributions of myosin II heavy chain B (by antibody immunofluorescence) (<i>a2, b2, c2</i>) and F-actin (by phalliodin_Alexa647 fluorescence) (<i>a3, b3, c3</i>). <b><i>a1</i></b>, Normal pattern of HB1.F3.eGFP::U251.dsRed cell interactions (inset shows U251.dsRed cell encirclement by a HB1.F3.eGFP cell). <b><i>a2, a3</i></b>, Normal distributions of MHC-IIB and F-actin. <b><i>b1</i></b>, Disruption of target cell encirclement after MHC-IIB inhibition by blebbistatin (50 µM; 18 h), contraction of MHC-IIB (<b><i>b2</i></b>) and formation of long actin-lined tubes emanating from both tumor cells and NSCs (<b><i>b3</i></b>). <b><i>c1</i></b>, Loss of lamellipodia and truncation of target cell encirclement (inset) after F-actin inhibition by cytochalasin D (1 µM; 18 h), and loss of filamentous structure and appearance of multiple aggregates by both MHC-IIB (<b><i>c2</i></b>) and F-actin (<b><i>c3</i></b>) and loss of spike-like filopodial structures emerging from both HB1.F3.eGFP and U251.dsRed cells (<b><i>c3</i></b>). <b><i>B</i></b>, <i>Above</i>, Proportions of HB1.F3.eGFP:: target cell pairs were not significantly affected by actin or myosin inhibition (<i>p</i>>0.05). <i>Below</i>, After actin disruption or myosin inhibition, encirclement was blocked and did not progress beyond type II (<i>p</i><0.05).</p