The requirement of G2A in J774A.1 cell chemotaxis to LPC. (A) Western blot of G2A expression in wild-type (WT), G2AsiRNA, and G2AHIGH J774A.1 cells. The lysate of G2AHIGH J774A.1 cells was diluted 10- and 100-fold before loading. (B) Comparison of LPC-induced chemotaxis of wild-type, G2AsiRNA, and G2AHIGH J774A.1 cells. Transwell migration assays were used to examine the role of G2A in J774A.1 cell chemotaxis to different concentrations of LPC. (C) As a control for cell motility, 5 nM C5a was used as a chemoattractant for J774A.1 cell migration under the same experimental condition. (D) Rescue of G2AsiRNA J774A.1 cell chemotaxis to LPC by reconstitution of G2A expression. An siRNA-resistant form of G2A cDNA was cloned into retroviral vector under the control of the ubiquitin C promoter, and G2AsiRNA J774A.1 cells were infected with this construct to reconstitute the expression of G2A. The chemotactic response to LPC of these cells (G2ARESCUE) was compared with wild-type, G2AsiRNA, and G2AHIGH J774A.1 cells. (E) Effects of different species of LPC on J774A.1 cell chemotaxis. LPC with different length of acyl chain was used as a chemoattractant to induce G2AHIGH J774A.1 cell migration. These results are representative of at least 3 independent experiments.
