Figure 2
Hsp90α knockout eliminates motility and invasiveness of the tumour cells. Serum-starved cells were subjected to colloidal gold motility (5000 cells per well) and the Matrigel invasion (20 000 cells per well) assays. (A) The migration tracks of MDA-MB-231, HBL-100 and human keratinocytes without (−) or with (+) 10% foetal bovine serum were quantitated by a computer-assisted analysis as MI (Materials and methods), as shown. (B) The motility measurements of the native MDA-MB-231, KO-α-#1 and KO-α-#2 cells in the absence (−) or presence (+) of human recombinant Hsp90α or human recombinant Hsp90β protein stimulation. (C) Matrigel invasion assay of the indicated cell lines with or without added human recombinant Hsp90α or human recombinant Hsp90β protein and the results quantitated as percentage of the invaded cells over the total number of seeded cells (Inv. %). (D) Evidence for FPLC-purified human recombinant Hsp90α and human recombinant Hsp90β proteins used in the above rescue experiments. (E) Evidence for downregulation of LRP-1 receptor in MDA-MB-231 cells by a lentivirus-delivered short hairpin RNA. (F) Matrigel invasion assay of LRP-1-downregulated cells in the absence or presence of human recombinant Hsp90α protein. Data were representative of multiple (n⩾ 4) independent experiments and represented as mean±s.e.m. P<0.05.
