Figure 1.

Hax1 expression in PLB-985 cells and localization during chemotaxis. (A) Schematic of Hax1 modified from (Jeyaraju et al., 2009). Binding sites for HS1 (Suzuki et al., 1997), Gα13 (Radhika et al., 2004), and β6 integrin (Ramsay et al., 2007) are shown. Hax1-GFP constructs are used in C. (B) Hax1 expression is decreased upon differentiation. Vinculin, HS1, and actin were included as controls and show increased expression with neutrophil differentiation. Quantification of expression represents the ratio of differentiated to undifferentiated cells as mean intensity ± SDs from three independent experiments as described in Materials and methods. (C) Still image from time-lapse videos of utrophin-RFP and Hax1-GFP localization in PLB-985 cells. Utrophin-RFP was coexpressed with full-length Hax1-GFP (Video 1), Hax1 1–113–GFP (Video 2), and Hax1Δ113-GFP (Video 3) in differentiated PLB-985 cells and then imaged during chemotaxis on 10 µg/ml fibrinogen in response to a needle containing 1 µM fMLP. Bar, 10 µm.