Extended Data Fig. 9: The curvature preference of integrin β5 is not reduced upon the knockdown of AP2-μ, clathrin heavy chain (CHC), EPS15/R, Intersectin1/2, but is significantly reduced upon the knockdown of FCHo1/2.

a, Representative fluorescence images showing that the shRNA knockdown of AP2-μ, clathrin heavy chain (CHC), EPS15/R, or Intersectin1/2 does not affect the accumulation of ITGβ5-GFP accumulation at the ends of vitronectin-coated nanobars in U2OS cells expressing RFP-CaaX membrane marker. BFP expression is a marker of shRNA transfection. Quantifications of the β5-GFP curvature preference under these conditions are presented in Fig. 3d. b, Immunofluorescence showing that the transfection of AP2-μ shRNAs (indicated by BFP expression) can reduce the appearance of AP2 complexes in U2OS cells and the accumulation of AP2 complex at the ends of vitronectin-coated nanobars. c, Left: Representative fluorescence images showing that knockdown of FCHo1/2 reduces the β5-GFP accumulation at vitronectin-coated nanobar ends. Right: Quantifications of the β5-GFP curvature preference in U2OS cells transfected with scramble or FCHo1/2 shRNAs. N = 12 cells for each condition with each cell represents the average of over 100 nanopillars. Data are mean ± SD. P values calculated using t-test. Scale bars: 10 μm.