Downregulation of TFPI in breast cancer cells induces tyrosine phosphorylation signaling and increases metastatic growth by stimulating cell motility

Table 2 Stable downregulation of TFPIβ in MDA-MB-231 breast cancer cells.

		TFPIβ/α mRNA downregulation, % of control	Total/Free TFPI ag	Total TFPI ag, PI-PLC/Untreated	Free TFPI ag, PI-PLC/Untreated
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.

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