Figure 2: Caveolin-1 accumulates in the rapidly retracting cellular edge during mitotic cell rounding. | Nature Communications

Figure 2: Caveolin-1 accumulates in the rapidly retracting cellular edge during mitotic cell rounding.

From: Interphase adhesion geometry is transmitted to an internal regulator for spindle orientation via caveolin-1

Figure 2

(a) Time-lapse serial ratio images of the RhoA FRET probe expressed in HeLa cells cultured on a fibronectin-line micropattern. (b) Images of a metaphase cell cultured on the line micropattern stained with anti-Gαi1 (green) and anti-caveolin-1 (red) antibodies. A middle section of Z-stack images (left) and an X-Z projection image in the boxed area (right) are shown. (c) Images of a metaphase cell expressing MyrPalm-GFP (green) cultured on the line pattern stained with an anti-caveolin-1 antibody (red). A middle section of Z-stack images (left), a Z projection image (middle), and an X-Z projection image in the boxed area (right) are shown. (d) Images of a mitotic cell cultured on a fibronectin-coated coverslip and stained with an anti-Gαi1 antibody, FITC-labelled cholera toxin subunit B (CTX-FITC), and Hoechst. (e) Images of GFP-caveolin-1 fluorescence and electron microscopy. The boxed area is enlarged (middle and right). Red dots and arrowheads represent caveola-like structures, and yellow dots and arrowheads represent caveosome-like structures. (f) Time-lapse images of a mitotic cell expressing GFP-LGN (green), RFP-caveolin-1 (red) and CFP-histone H1 (blue) on the line micropattern. Arrowheads indicate the recruitment of fluorescent proteins at the retracting cortical region. (g) Angular distributions of the spindle orientation along the long axis of the line pattern. (h) The distribution of cortical caveolin-1 or Gαi1 signals against the spindle axis (white). The colour-code indicates the average of the relative intensity at each angle. (i) Distribution of the angle difference between the spindle angle and predicted spindle orientation based on the cortical intensity of caveolin-1 or Gαi1. P values represent the probability against random prediction (within 30°). Mean±s.d. (below). (j) Distribution of retraction velocity per angle (left) and distribution of values per angle at the cellular edge (right). For details, see Supplementary Figs 3 and 4.

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