Dual Role of 64 Integrin in Epidermal Tumor Growth: Tumor-suppressive Versus Tumor-promoting Function

Supplemental Material

This article contains the following supporting material:

• Supplementary Figure 1 - Molecular and growth characteristics of mTIC variants. (A) Ras pull-down assay using the Ras binding domain of Raf fused to GST. The levels of active Ras in mTICs are comparable to those in the immortalized NMK-1 line, but they are much lower than in the Ras-transformed mSCC1 line. Activation of Ras in the mTIC line can be triggered by EGF stimulation, indicating that Ras is not constitutively activated by mutation. (B) TOP/FOP luciferase assay showing that mTICs have a TOP/FOP ratio of around 1 reflecting a background transactivating activity of β-catenin/Tcf. The co-expression of a constitutively active form of TCA (TCA^CA) increases the TOP/FOP ratio. The * indicates that the reporter activity is significantly different from the control situation (p<0.05). (C) Immunofluorescent analysis of β-catenin in mTICs and mTIC-derived tumors shows no specific nuclear staining, suggesting that the β-catenin/Tcf pathway is not constitutively activated. Bar, 10 μm. (D,E) Growth curves of the tumor derived from mTICs (D) and mTICs-Ras^V12 (E) injections. This graph shows one representative experiment each of 6 injections (3 mice). The volume of the tumors was measured in 2 directions (L, length and W, width) and volumes are determined by (L x W x ((L+W)/2)).