Laminin-332 expression in stromal fibroblasts is upregulated by invasive breast cancer cells. (A) Isolation of stromal fibroblasts from invasive ductal carcinoma (IDC). Based on the zonal concept introduced in our previous study, fibroblasts were isolated from early-stage IDC tissue (less than 10 mm diameter) by enzyme digestion after confirmation of laminin-γ2 expression. (B) Expression of laminin-γ2 and α-SMA in fibroblasts of IDC. (C) Stimulation of laminin-γ2 expression in fibroblasts by CM from cancer cells. Fibroblasts were incubated with RSM, MCF7 CM, or MDA-MB-231 CM for 24 hours. (D) Duration of laminin-γ2 expression in interface fibroblasts (InFs) under anoikis conditions. InFs (5 × 105 cell/well) were seeded onto poly-HEMA coated six-well plates with RSM or MDA-MB-231 CM for 72 hours. Suspended cells were collected at 6, 24, 48, and 72 hours, as indicated.