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. 2014 Nov 7;125(4):649–657. doi: 10.1182/blood-2014-04-571331

Figure 7.

Figure 7

HGAL palmitoylation and myristoylation mutants enhance RhoA activation and greatly inhibit cell motility. (A) RCK8 cells stably expressing WT and G2A, C43A/C45A, and G2A/C43A/C45A HGAL mutants were starved for 8 hours and then seeded on fibronectin for 60 minutes. Cellular extracts were prepared and RhoA pull-down assay was performed. Equal loading was confirmed by immunoblotting with actin antibodies. Results are representative of 3 independent experiments. Densitometry analysis of normalized RhoA-GTP to total RhoA is presented. (B) RCK8 cells stably expressing mock vector or WT, G2A, C43A/C45A, and G2A/C43A/C45A HGAL mutants were used for IL-6 or SDF-1 chemotaxis assay performed in triplicate. Data are expressed as the mean ± standard error of the mean of triplicates. *P < .05; **P < .001. Results are representative of 2 independent experiments. GTP, guanosine triphosphate.