Fig. 7: EHD2, pacsin2 and EHBP1 stabilize caveolae at the plasma membrane.

a Representative PREM example images of wildtype MEFs (wt), EHD2 knockout MEFs (KO), pacsin2 siRNA or EHBP1 siRNA treated MEFs. Triple knockout/knockdown (KO/KD) indicates EHD2 KO MEFs that were treated with pacsin2 and EHBP1 siRNA. Scale bar is 200 nm. b Example PREM image of low, medium and highly curved caveolae. c In PREM images the total caveolae number at the plasma membrane in wt, EHD2 KO, pacsin2 siRNA treated, EHBP1 siRNA treated, or triple KO/KD MEFs was measured (n(wt) = 25 cell regions, n(EHD2 KO) = 13 cell regions, n(pacsin2 siRNA) = 16 cell regions, n(EHBP1 siRNA) = 15 cell regions, n(Triple KO/KD) = 16 cell regions, 3 independent experiments). d Number of individual caveolae types in wt, EHD2 KO, pacsin2 siRNA, EHBP1 siRNA or triple KO/KOD MEFs (n(wt) = 13 cell regions, n(EHD2 KO) = 13 cell regions, n(pacsin2 siRNA) = 17 cell regions, n(EHBP1 siRNA) = 15 cell regions, n(Triple KO/KD) = 16 cell regions, 3 independent experiments). e Caveolae radius (round caveolae domains were assumed) of low, medium and highly curved caveolae in wt, EHD2 KO, pacsin2 siRNA, EHBP1 siRNA or triple KO/KOD MEFs (caveolae number: low: n(wt) = 127, n(EHD2 KO) = 139, n(pacsin2 siRNA) = 163, n(EHBP1 siRNA) = 123, n(Triple KO/KD) = 69; medium: n(wt) = 148, n(EHD2 KO) = 130, n(pacsin2 siRNA) = 146, n(EHBP1 siRNA) = 177, n(Triple KO/KD) = 145; high: n(wt) = 129, n(EHD2 KO) = 157, n(pacsin2 siRNA) = 106, n(EHBP1 siRNA) = 142, n(Triple KO/KD) = 133, 3 independent experiments). Box plots indicate mean values ± SE, whiskers show SD, each replicate is depicted, statistical significance was measured by two-sided t test in normally distributed data sets, otherwise the nonparametric two-sided Mann–Whitney test was applied.