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. Author manuscript; available in PMC: 2014 Dec 1.
Published in final edited form as: Clin Cancer Res. 2013 Oct 11;19(23):6461–6472. doi: 10.1158/1078-0432.CCR-13-1284

Figure 2.

Figure 2

Effects of ATX and its catalytically inactive mutant (T210A) on RCC and endothelial cells. A, HRC-223 (RCC) and HUVECs were serum-starved for 4 hours and treated with conditioned media containing ATX or its mutant for 30 minutes. Cell lysates were collected and analyzed by immunoblotting using the indicated antibodies. B, HRC-223 and HUVECs were seeded on E-Plates at 10,000 cells per well and continuously monitored for impedance using The xCELLigence System. Arrowhead indicates the time point at which conditioned media from HEK293 cells transfected with vectors encoding the indicated proteins were added. C, HRC-223 and HUVECs were assessed for their chemotactic response to conditioned media containing ATX, T210A, or ATX plus pertussis toxin (PTX) using a modified Boyden chamber assay. The assays were performed in triplicate and quantified under light microscopy by counting three randomly chosen HPFs for each replicate (Student’s t-test; *, P<0.01).