Figure 7. Flagella are sufficient to phosphorylate EGFR and Shc.

Calu-3 cells were grown as well polarized monolayers on Transwells for 9 days and treated with heparinase III (hepIII), tunicamycin (tun), or EGFR inhibitor (AG1478). As a control, cell were left untreated (un); flagella-, Tfp-, or, as a negative control, BSA-coated beads were added to the AP or BL chamber for 1 h. Lysates were immunoprecipitated with Akt or EGFR antibody or directly immunoblotted with (A, B) phospho-Akt, (C–F) phospho-EGFR, or (G–H) phospho-Shc (three different isoforms p46, p52, and p66). Representative gels (A, C, E, G) and quantification by densitometry of three gels (B, D, F, H) are shown. The ratio of phospho-Akt to total-Akt, phospho-EGFR to total-EGFR, or phospho-Shc to total-Shc for untreated cells was set to 1. Shown is the mean +/− SD for 3 independent experiments. ^**P<0.05 compared to cells BL infected with PAO1.