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. 2014 Nov 5;289(52):35757–35769. doi: 10.1074/jbc.M114.595512

FIGURE 5.

FIGURE 5.

LPA-induced macromolecular complex formation of LPA2 on the plasma membrane. A, co-immunoprecipitation was performed to test the macromolecular complex formation of LPA2, NHERF2, and PLC-β3. B, the bar graph depicts the average ratios of PLC-β3 and NHERF2 with LPA treatment to their corresponding controls (without LPA treatment). The data are densitometry readings from the immunoreactive bands as represented in A (mean ± S.E. (error bars), n = 3). C, MSD versus time plots for LPA2 in 3T3-LPA2 cells with or without knockdown of NHERF2 expression and in the presence or absence of LPA. The inset blots show the knocking down of NHERF2 expression in 3T3-LPA2 cells. D, MSD versus time plots of LPA2 in 3T3-LPA2 cells treated with 1 μm LatB for 30 min in the absence or presence of 1 μm LPA. E, MSD versus time plots of LPA2 in 3T3-LPA2 cells treated with 30 μm nocodazole for 30 min in the absence or presence of 1 μm LPA. F, the kinetics of the relative wound density from wound healing assays conducted in the IncuCyteTM high content imaging system. The assays were performed using DKO-LPA2 MEFs with or without transient knockdown of NHERF2 expression. The inset blots show the knocking down of NHERF2 expression in DKO-LPA2 MEFs. G, quantitative analysis of cells migration as shown in F. IB, immunoblot.