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. Author manuscript; available in PMC: 2012 Mar 1.
Published in final edited form as: Gastroenterology. 2010 Dec 4;140(3):924–934. doi: 10.1053/j.gastro.2010.11.054

Figure 2. MAGI-3 competes with NHERF-2 for interaction with LPA2 to attenuate PLC activity.

Figure 2

(A) HCT116 cells stably expressing VSVG-LPA2 (HCT116/LPA2) were transfected with MAGI-3 siRNA or MAGI-3. Transfected cells were treated with 1 μM LPA for 20 min, NHERF-2 was immunoprecipitated, and co-immunoprecipitated VSVG-LPA2 was detected (top two panels). The bottom two panels show LPA2 and MAGI-3 in cell lysates. (B) The interaction between LPA2 and MAGI-3 in HCT116/LPA2 cells transfected with NHERF-2 siRNA or NHERF-2 was determined as described above. The top two panels show co-immunoprecipitated LPA2 and immunoprecipitated MAGI-3. The expression of LPA2 and NHERF-2 in cell lysate is shown in the bottom panels. (C) The PLC activation by LPA in HCT116/pcDNA or HCT116/NHERF-2 cells was determined as described in Methods. The data are represented as relative percent change compared with respective untreated cells. n=3. The amounts of IPs generated by LPA were determined in (D) HCT116 cells overexpressing MAGI-3 and (E) MAGI-3 knockdown cells. (F) The interaction between V5-MAGI-3 and Flag-PLCβ was determined. PLCβ expression is shown in the right panel. n ≥ 3 for each experimental set.