Figure 5.

Defective Polarization and p-HS1 Accumulation in B Cells Lacking the ABD of WIP

(A) Representative SIM images (maximum intensity projections) of phalloidin and p-HS1 staining in fixed WT, Wipf1^−/−, and WIPΔABD B cells settled on coverslips coated with poly-L-lysin and treated with CXCL12 (500 ng/mL) for 3 min. Scale bar, 4 μm.

(B and C) Quantifications show the percentage (mean ± SEM) of polarized cells (each dot represents an image with about 50 cells each) after stimulation with (B) 500 ng/mL or (C) 200 ng/mL CXCL12.

(D–F) Quantification of the (D) polarized area, (E) MFI of p-HS1 per cell, and (F) fluorescence intensity of p-HS1 in the polarized area as analyzed by the ImageJ software. Each dot represents a cell. About 150 cells were analyzed per genotype. Data (mean ± SEM) are representative of at least two independent experiments.

(G) Immunoblot of splenic WT and WIPΔABD B cells treated with CXCL12 and probed with antibodies against p-HS1, HS1, and tubulin. Data (mean ± SEM) are representative of two independent experiments.

See also Figure S3.