(A) PLCD1-expressing cells and vector-only control cells (MDA-MB-231 and MCF-7) and BT-549 cells transfected with siN.C. and siPLCD1 for 72 h. PLCD1, pERK1/2, ERK1/2, pGSK-3β, GSK-3β, active-β-catenin, β-catenin, and MMP7 protein levels were measured by immunoblotting with β-actin as an internal control, and supernatant MMP7 levels were also measured by ELISA. Data were based on at least three independent assays, and representative images were shown on the left. Means ±SD, **p<0.01. (B) Expression of related proteins was detected by immunoblotting at the indicated time points (0 h, 1 h, 12 h, 24 h) after treatment with U0126 in MDA-MB-231 and MCF-7 cells. Supernatant MMP7 levels were also measured by ELISA. Data were based on at least three independent assays, and representative images were shown. Means ±SD, **p<0.01. (C) Cells transfected with siN.C. or siPLCD1 were treated with or without U0126 for 1 h, and protein levels were measured by immunoblotting in BT-549 cells. Supernatant MMP7 levels were also measured by ELISA. Data were based on at least three independent assays, and representative images were shown. Means ±SD, **p<0.01.