Fig. 2: Expansion Microscopy unveils the periodic organization of spectrin clusters. | Nature Communications

Fig. 2: Expansion Microscopy unveils the periodic organization of spectrin clusters.

From: Mechanically induced topological transition of spectrin regulates its distribution in the mammalian cell cortex

Fig. 2

A Cartoon representation of actin-spectrin tetramers (estimated length ≈ 180/190 nm) and the relative positions of the immuno-reactive epitopes (Y symbols). B The experimental pipeline to enhance the spatial resolution by Expansion Microscopy (ExM). C Representative ExM images of MEFs immunolabelled for βII-spectrin (green) and β-actin (magenta) are shown; in white is reported the “real” scale bar of the image (10 μm) and in red the extrapolated scale according to the expansion factor of 4x. The zooms related to βII-spectrin white boxes (1–6) are shown to highlight periodic clusters (D), and the corresponding intensity line scans and peak-to-peak distances are reported (real in white and extrapolated in red). E Representative ExM image showing βII-spectrin (green) in the diffused (zoom 1, white dashed box) and constrained mesh configurations between stress fibers are shown (zoom 2). Images are representative of at least 3 independent experiments. F Nearest Neighbor Distance distributions in the two configurations are calculated for the βII-spectrin epitopes. Adjacent zones within the same cells can display large discrepancies in terms of spectrin organizations (drawn in G). Panel (B) was created with BioRender.com and released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license.

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