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Fig. 7. Internalised MT1-MMP colocalised with caveolin-1 and clathrin heavy chain
and is present in caveolin-1 containing fractions. bbHT1080 cells, which had
internalised IgG-bound MT1-MMP complexes for 10 minutes (A,D), were fixed,
permeabilised and incubated with antibodies directed against caveolin-1 (B)
and clathrin heavy chain (E) followed by a fluorescently labelled secondary
antibody. Merged panels (C,F) are shown in the right-hand column. Arrowheads
are examples of colocalising structures. Bar, 20 µm. (G) After a
detergent-free carbonate-based extraction, HT1080 cells were homogenised and
subjected to subcellular fractionation. Fractions (1 ml) were collected from
the top of the gradient, separated on a 10% SDS-PAGE gel and immunoblotted
using anti-MT1-MMP affinity-purified IgG (upper panel) or with an
anti-caveolin-1 pAb (lower panel).