Images from fixed tissues were acquired using an LSM 700 microscope (Zeiss) equipped with a ×63/1.4 Plan Apochromat DIC oil immersion objective. Quantifications of pMLC2 fluorescence intensities at the back, the front, and the side of the cluster were performed as previously described6 (link).
For more precise analysis of pMLC2 localization and for the line scan analysis, we acquired images from fixed tissues with the Airyscan detector of a Zeiss LSM880 confocal equipped with a ×63/1.4 Plan Apochromat oil immersion objective. Line scans were performed on the Zen Blue software and plotted in GraphPad Prism. Specific line scan were chosen according to the base of the protrusion. The protrusion bases were determined manually by its neck shape (opposite negative curvature shapes). The signal were normalized to the maximal signal of pSqh intensity.
For more precise analysis of pMLC2 localization and for the line scan analysis, we acquired images from fixed tissues with the Airyscan detector of a Zeiss LSM880 confocal equipped with a ×63/1.4 Plan Apochromat oil immersion objective. Line scans were performed on the Zen Blue software and plotted in GraphPad Prism. Specific line scan were chosen according to the base of the protrusion. The protrusion bases were determined manually by its neck shape (opposite negative curvature shapes). The signal were normalized to the maximal signal of pSqh intensity.
Free full text: Click here
