Figure 2. Endocytic membrane morphologies in chc1Δ cells.

(A) Correlative fluorescence and electron microscopy of chc1Δ yeast cells expressing Sla1-GFP, Abp1-mCherry. Three examples of membrane ultrastructures underlying sites of Sla1 and Abp1 colocalization. Upper panel shows overlays of RFP and GFP channel images, lower panel images are virtual slices through electron tomograms, acquired at positions corresponding to the fluorescent spots marked by the white dashed circles. Black arrowheads indicate endocytic vesicles. Scale bars are 2 µm in the upper panel, 100 nm in the lower panel. (B) Curvature of invagination tips identified by correlative microscopy in chc1Δ, plotted against the invagination depths. The dashed line is a cubic smoothing spline fitted to the data points of wild-type invagination tip curvatures, published in (Kukulski et al., 2012a). (C) Surface areas of vesicles identified by correlative microscopy in chc1Δ and compared to wild-type cells (Kukulski et al., 2012a). Red central line represents the mean, upper and lower red lines represent standard deviations.

DOI: http://dx.doi.org/10.7554/eLife.16036.004

Figure 2—figure supplement 1. Membrane shape parameters in chc1Δ cells.

(A) Appearance of tubular segments. The angles between the two sides of invagination membrane profiles are plotted against invagination depth. When the angle reaches zero degrees, the invagination consists of a tubular segment. Wild-type data points are from (Kukulski et al., 2012a). The dashed lines show the regression line describing the wild-type data and its prediction band (95% prediction interval), calculated based on invaginations with a membrane angle >0. The difference between the wild-type and chc1Δ data points is not significant (p value > 0.1). (B) Position of the neck along the invagination. Only invaginations with a width that corresponds to less than 80% of the tip diameter are considered to have a neck. The distance of the neck to the invagination tip is plotted against invagination depth. Wild-type data points are from (Kukulski et al., 2012a). The percentage of invaginations with neck (19% of all chc1Δ invaginations have a neck) is similar to wild-type cells (27% of all wild-type invaginations have a neck).

DOI: http://dx.doi.org/10.7554/eLife.16036.005