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. Author manuscript; available in PMC: 2012 Sep 1.
Published in final edited form as: Cancer Res. 2011 Jul 20;71(17):5915–5922. doi: 10.1158/0008-5472.CAN-11-0466

Figure 2.

Figure 2

Mesothelin shedding is inhibited by broad-spectrum MMP/ADAM inhibitors. A. A431/H9 cells harvested by trypsinization were incubated with PI-PLC (1 U/mL) or PC-PLC (10 U/mL) for 1 h at 37°C. After wash with FACS buffer, cells were stained by Alexa488-labeled SS1P (2 μg/mL). The fluorescence signal was measured by flow cytometer. Cells without SS1P-Alexa488 staining were used as the negative control. Cells incubated with Hank’s buffer and stained by SS1P-Alexa488 were used as positive control. B and C. A431/H9 cells were seeded into 96-well plate and incubated with phospholipase inhibitor (B) or pan-MMP/ADAM inhibitors (C), including U73122 (10 μM), ET-18-OCH3 (10 μM), sPLA2-IIA inhibitor I (50 μg/mL), D609 (250 μg/mL), neomycin (1 mg/mL), GM6001 (20 μM), batimastat (20 μM), marimastat (20 μM) and camostat (20 μM). After 48 h, mesothelin in cell medium was measured by ELISA. DMSO-treated cell was negative control. The results were normalized with cell number in each well.