Ghest level of vimentin and perks. The expression of EGFR and ERK2 in Figure 4 Cell 1 Versican G3 domain structure and variable expression in mouse mammary tumor cells, versican. Immunoblotting showed that 4T1 cells, the h HIGHEST versican V1 isoform, vimentin, cells Perk, 67NR and 66c14 expressed N-cadherin expressed, w During 4Q07 and 4T1 cells express ABT-492 inhibitor E-cadherin, 20 ng / ml EGF medium, expressed 4T1 cells obtained ht pEGFR and perks. EGFR *: addition of 20 ng / ml EGF for 5 min, ERK *: addition of 20 ng / ml EGF for 60 minutes. Versican G3 construct was expressed in cells analyzed by Western blot 66c14 with cell lysate and culture medium. Appeared morphologically the G3-transfected cells 66c14 stretched more and uniformly Strength distributed over the substantially rectangular cells in the control group that had a tendency to aggregate in clusters.
doi: 10.1371/journal.pone.0013828.g001 AMG 900 945595-80-2 vascular versican promotes EGFR signals PLoS ONE | www.plosone fourth November 2010 | Volume 5 | Issue 11 | e13828 lines was similar. 67NR and 66c14 cells expressed N-cadherin, w During 4Q07 and 4T1 cells express E-cadherin. If with 20 ng / ml EGF treated for 5 minutes, expressed 4T1 cells, the h HIGHEST EGFR p. When 4T1 cells were treated with 20 ng / ml EGF increased for 60 minutes Hte observed expression of pERK. To investigate the effect of versican G3 on the growth and metastasis of breast cancer cells, and its potential signaling pathways, we have exogenously GE U Versican G3 construct ert 66c14 cells.
The expression of versican G3 transfected into cell lysate and culture media of cells 66c14 cells is compared with the vector control also shown in Figure 1b. Morphologically, cells transfected 66c14 G3 seemed more l Accessible and equitable distribution of in vitro compared to the predominant aspect of the cuboid Shaped cells tended to aggregate in groups in the vector control group. Versican G3 enhances cell adhesion Sion of breast cancer in studies of transfected cell binding, G3-and vector-66c14, 4Q07-4T1 cells and the cells were seeded in 6 bo Their culture. After the cells were incubated in 2.5% FBS / DMEM for 2 hours, we observed increased Hte cell attachment to culture dishes in G3 as compared to the thwart of vectors. Growing area 2.5, 5 and 10% FBS / DMEM for 3 hours was observed that most G3 66c14 cells transfected of food.
Block EGFR with AG 1478, or treating the cells with selective MEK-inhibitor PD 98059 had no effect on the G3-induced cell attachment reviewed in time. Versican G3 activates the path of the EGFR / ERK immunoblot showed that the expression construct into cells G3 66c14 MODIFIED not alter the total protein of EGFR, ERK2, and N-cadherin, but increased Ht fa is spectacular r levels of pEGFR and perks. The presence of G3 also regulated expression of fibronectin and down-regulated expression of vimentin. Grown in 20 ng / ml EGF for 5 middle of the 60 minutes suppressed the G3-transfected cells, increased levels of pEGFR ht And perks. With 20 ng / ml EGF and various concentrations of EGFR antagonist AG 1478, be k Nnte G3 activated pEGFR with increasing dose blocked by inhibitors. PERK expression was also inhibited in cultured cells, the G3 in medium with 5.0 mm AG 1478th With 20 ng / ml EGF and various concentrations of selective MEK-inhibitor PD 98059, G3-induced expression of pERK, pEGFR but not, k nnte Be blocked by PD 98059. Versican G3 term