|
|
|
|
|
|
|
|
|||||
| Home Help Feedback Subscriptions Archive Search Table of Contents | |||||
|
Fig. 5. S100A4 acts as a crucial downstream effecter of CTGF. (A) cDNA microarray analysis (Cy3-Cy5 merged image) showing the expression of S100A4, arrow indicates human S100A4. (B) Expression of S100A4 mRNA and protein by RT-PCR and western blotting, respectively. (C) Effects of ERK1/2 activation on CTGF-regulated S100A4 expression. MCF-7/neo and MCF-7/CTGF cells were treated with DMSO or PD98059 (20 µM) for 24 hours, and the levels of S100A4 were evaluated by western blotting. (D) MDA231/neo and MDA231/AS cells were transiently transfected by constitutively activated MEK1; 48 hours after transfection, expression levels of S100A4 were evaluated by western blotting. (E) Effects of S100A4 on CTGF-mediated cellular motility. MCF-7/neo and MCF-7/CTGF cells were stably transfected with pcDNA4 or pcDNA4-AS-S100A4, respectively (see Materials and Methods). Expression levels of S100A4 were detected by western blotting (upper panel). Cellular motility was measured by migration assay (lower panel). (F) MDA231/neo and MDA231/AS cells were stably transfected with pcDNA4 or pcDNA4-S100A4, respectively. Expression levels of S100A4 were detected by western blotting (upper panel). Cellular motility was measured by migration assay (lower panel). (G) S100A4 is involved in CTGF-mediated cytoskeletal changes. Cells were fixed in 3.7% paraformaldehyde and photographed under a light microscope (a,d,g,i). The fixed cells were co-stained with Texas-Red-phalloidin for F-actin (red; b,e,h,k) and monoclonal antibody against paxillin (green; c,f,i,l). (H) S100A4 expression is required for CTGF-mediated metastatic colonization. Lungs of female BALE/cAnN-Foxn1nu/CrlNarl nude mice were excised and photographed after the experimental metastasis assay.
|
