16HBE cells were stably infected with pSUPER, pSUPER expressing shCdc42, shARHGEF18, shSOS1 hairpins 1-3, pQCXIP or pQCXIP expressing dominant-negative myc-HRas N17.
Cells were seeded sparsely on glass coverslips, incubated for 3 days and then fixed and stained for ZO-1 (tight junctions) and DNA (nuclei). Scale bar, 20 μm.
Cell lysates were analysed by Western blotting for SOS1 and actin; labels as for (C).
Quantification of tight junction phenotype. > 500 cells were counted per sample/experiment, across n = 3 independent experiments (dots indicate individual data points). Error bars denote mean ± SEM. *P = 0.0199; **P = 0.0041; ****P = 0.0001.
Cell lysates were analysed by Western blotting for Ras, pERK and total ERK.
Cells were seeded sparsely on glass coverslips, incubated for 3 days and then fixed and stained for ZO-1 (tight junctions) and DNA. Scale bar, 20 μm.
Quantification of tight junction phenotype (see C). ***P = 0.0002.
Data information: All data are representative of 3 independent experiments.