Fig. 6: Spatially distribution of EHD2, pacsin2 and EHBP1.

a Representative STED-CLEM images for MEFs expressing EHD2-EGFP (labelled with GFP nanobody-Atto647N) or white arrows indicate accumulation of EHD2 around the caveolae center. b EHD2 STED fluorescence profile from the center of caveolae to its edges obtained from STED-CLEM images (a). Each individual caveolae type is depicted (graph shows mean ± SE, caveolae number: n(low) = 72, n(medium) = 50, n(high) = 163, 2 independent experiments). c STED-CLEM showing endogenous EHD2 antibody staining at low curved caveolae (secondary antibody tagged with Atto647N, 2 independent experiments). d,e Representative CLEM images for MEFs expressing either pacsin2-EGFP (d) or EHBP1-EGFP (e) that were labelled with GFP nanobody-Atto647N (cyan). f Quantitative analysis of low or highly curved caveolae by STED fluorescence profiles from the center of caveolae to their edges (indicated by green dashed line, line graphs show mean ± SE, caveolae number: caveolin1: n(low) = 145, n(high) = 178; EHD2: n(low) = 72, n(high) = 163; pacsin2: n(low) = 103, n(high) = 77; EHBP1: n(low) = 70, n(high) = 79, 3 independent experiments). g Percentage of low or highly curved caveolae that were targeted by either pacsin2 or EHBP1. Bar plot indicates mean ± SE, n(pacsin2) = 1357 caveolae/3 cells, n(EHBP1) = 660 caveolae/4 cells, 3 independent experiments. Significant difference was tested by two-sided Mann–Whitney test. Scale bar is 100 nm.