Fig. 1 | Nature Communications

Fig. 1

From: Superresolution architecture of cornerstone focal adhesions in human pluripotent stem cells

Fig. 1

Characterisation of cornerstone adhesions. a Spinning disk images of hPSC plated on vitronectin (VTN) and stained for F-actin, paxillin, DAPI, and the pluripotency marker Nanog. Scale bar 10 μm. b Structured illumination microscopy images of hPSC plated on VTN and stained for filamentous actin (F-actin) and paxillin. Scale bar 10 μm. The white square highlights a region of interest (ROI), which is magnified. c–f Live-cell imaging of endogenously tagged paxillin in hPSC. Images were acquired every minute using a spinning disk microscope. Colonies were recorded for at least 105 min (n = 16 independent colonies from three biologically independent experiments). A representative video is available as supplementary information (Supplementary Movie 1). c A representative image is displayed (ROI highlighted by white square and magnified; size, 30 µm) in addition to a mask image highlighting paxillin-positive adhesions (colony edge FA, green; colony centre FA, magenta) and merged images depicting paxillin-positive adhesion lifetime (selected time points 0 min, green; 50 min, cyan and 100 min, magenta) within one hPSC colony (white represents very stable adhesions). d The percentage area covered by paxillin-positive adhesions at the edge or at the centre of hPSC colonies was measured over the duration of the movies. Statistical significance was determined by Student’s t-test (two-tailed, unpaired). e, f The frequency distributions of paxillin-positive adhesion lifetime (e) and maximal size (f) (colony edge FA, green circle; colony centre FA, magenta cross) within hPSC colonies are displayed. Each data point represents the frequency distributions in one hPSC colony. Statistics: multiple t-tests with correction using Holm–Sidak method. Error bars depict standard deviation. Source data are provided as a Source Data file

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