![]() You can choose the Grid view or List View. Data represent mean ± SD of triplicate samples. When you enter the Photo, Video, Music or File/Folder directory, all files in the specific directory will be listed. Confocal imaging and analysis were performed blind to the treatment conditions using a Leica SP8 confocal microscope and Leica Application Suite Software. The results shown are from one experiment, representative of three independent experiments. The relative MFI of CCR7 expression is shown (right). The gray-filled histogram represents staining with isotype control antibody (left). ( E) CCR7 expression levels were examined in the presence (red open histogram 2 mM, green open histogram 1 mM) or absence (blue open histogram) of MβCD by flow cytometric analysis using anti-human CCR7 antibody. Data represent mean ± SD of triplicate wells. The indicated chemokine was added to the lower wells, and H9 cells were added to the upper wells in the presence of the indicated concentrations of MβCD. ( D) Chemokine-induced cell migration with or without MβCD was examined using the Transwell assay. The results shown are from one experiment, representative of three independent experiments, with the mean number of the signals plotted on the vertical axis. The number of PLA signals per cell was counted using the Duolink Image Tool software. ![]() ( C) CCR7 homodimer formation after treatment with or without 2 mM MβCD was examined by in situ PLA (Red PLA signal, Blue Hoechst 33342). White arrowheads show CCR7 homodimers detected in the GM3-enriched lipid rafts. The center image illustrates a cross-sectional image (XY-plane), whereas the images to the right and below illustrate those in the YZ-plane and XZ-plane, respectively. ( B) Orthographic projection of the cell shown in ( A). During migration, cells were fixed and stained with anti-human CCR7 antibody conjugated with the complementary oligonucleotide probes, anti-GM3 antibody, and Hoechst 33342. After cell alignment was complete, human CCL21 (100 ng) was applied to the contra-wells. T cells were loaded into each well of the EZ-TAXIScan microchamber. ( A) The localization of CCR7 homodimers in T cell migration in response to CCL21 is shown (Red PLA signal, Green anti-GM3 antibody, Blue Hoechst 33342). CCR7 homodimers are polarized toward GM3-enriched leading edge during CCR7-dependent cell migration.
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