Assembly assay identifies a critical region of human fibrillin-1 required for 10–12 nm diameter microfibril biogenesis
Fig 4
Microfibril incorporation of GFP-FBN1 nMFS variants.
FS2 fibroblasts were co-cultured for 5 days with HEK293T cells transiently transfected to express GFP-FBN1 (WT) or variants associated with nMFS. Co-cultures were then fixed and stained with anti-GFP and anti-FBN1 antibodies without permeabilisation [16, 26]. Although the nMFS variants I1048T, E1073K, C1111R, N1131Y and ΔcbEGF16 were clearly detectable in medium samples when expressed as GFP-FBN1 fusions, microfibril networks labelled with the recombinant GFP-FBN1 (arrow in WT panel) were rarely seen. Bar = 100 μm.