|  | TFPIβ/α mRNA downregulation, % of control | Total/Free TFPI ag | Total TFPI ag, PI-PLC/Untreated | Free TFPI ag, PI-PLC/Untreated |
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MDA-MB-231 | pSiRPG-shRNA7β | 46/2 | 2.0/1.6 | 0.7/0.2 | 0.1/0.1 |
 | pSiRPG-shRNA9β | 42/1 | 2.3/1.6 | 0.8/0.2 | 0.1/0.1 |
 | pSiRPG (control) | 0/0 | 3.5/1.7 | 1.5/0.3 | 0.2/0.2 |
- MDA-MB-231 breast cancer cells were transduced with vectors containing shRNAs against TFPIβ or empty vector pSiRPG as a control. Relative TFPIα and β mRNA levels were quantified using qRT-PCR and cell-associated protein (ag) with total and free TFPI ELISA kits. GPI attached TFPI was measured in the supernatant after PI-PLC treatment using total and free TFPI ELISA kit. Ag levels were corrected against cellular total protein and presented as ng TFPI/mg total protein.