Establishment of stably transduced <i>Arhgef11</i> knockdown cell lines using shRNA lentiviral methodology.

<p>A number of shRNA lentiviral constructs (shRNA1-4) were tested for optimal knockdown of <i>Arhgef11</i> in both NRK and HEK293T cells (see <b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0132553#pone.0132553.s002" target="_blank">S2 Fig</a></b>). (<b>A</b>) Real-time PCR confirmation of shRNA3 <i>Arhgef11</i> knockdown and subsequent impact on RhoA and ROCK in NRK stably transduced cells compared to scrambled control (LVC). (<b>B</b>) Real-time PCR confirmation of shRNA3 <i>Arhgef11</i> knockdown and impact on RhoA and ROCK in HEK293T stably transduced cells compared to scrambled control (LVC). (<b>C</b>) Western analysis and densitometry measurements of ARHGEF11, RhoA, and ROCK between NRK-LVC and NRK-shRNA3.(<b>D</b>) Western analysis and densitometry measurements of ARHGEF11, RhoA, and ROCK between HEK293T-LVC and HEK293T-shRNA3. n = 4–6 independent samples/group, *p<0.05 versus LVC, Error bar are ±SD.