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. 2019 Mar 21;30(7):838–850. doi: 10.1091/mbc.E18-10-0652

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© 2019 Wang et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology.

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FIGURE 6: — Integrin β4E-tGFP distributes into ringlike patterns containing F-actin and CD9 at the cell–ECM surface. (A) Schematic shows the use of confluent cultures. (B) Epifluorescence images at the cell–ECM show the distribution of β4E-tGFP (green, B4E-tGTP), β4C (B4-CT, red), nuclei (blue), and the corresponding black-and-white split-channel images. Scale bar: 10 microns. (C) Epifluorescence images at the cell–ECM show the distribution of β4E-tGFP (green, B4E-tGTP), F-actin (Phalloidin, red), and nuclei (blue). Boxed region (C, far-left panel) at higher magnification (C, second panel) (scale bar: 10 microns) shows the distribution of β4E-tGFP (green, B4E-tGTP), F-actin (Phalloidin, red), and nuclei (blue), with the corresponding colocalization of white pixels map (C, third panel) and a merged image (C, far-right panel). x,z-projection (C′) shows β4E-tGFP localized at the cell–ECM plane. (D) Epifluorescence images (D, far-left panel) at the cell–ECM show the distribution of β4E-tGFP (green, B4E-tGTP), CD9 (CD9, red), and nuclei (blue). (D, second panel) shows distribution of CD9 (green), F-actin (phalloidin, red), and nuclei (blue). (E) Representative 3D-SIM superresolution images at different z-sections show the distribution of β4E-tGFP (green, B4E-tGTP) and F-actin (Phalloidin, red) as a ringlike pattern from cell–ECM to cytoplasm. Scale bars: 1 micron. (F) Representative images show β4C localized at HD-like structures at cell–ECM. Left panel shows the distribution of BP180 (Green, BP180) and β4C (red, B4C) at cell–ECM. Middle and right panels show the corresponding colocalization white pixels map between BP180 and β4C. HD-like structures were defined as colocalization of BP180 and β4C. Nuclei are shown in blue. Scale bar: 10 microns.