GEF-H1 knockdown inhibits complement-mediated RhoA activation and augments cell injury in GEC.
A, GEC were transduced with lentivirus encoding GEF-H1 shRNA or empty vector pLKO.1 (control). Seventy-two hours post-transduction, cell lysates were immunoblotted for GEF-H1. **, p < 0.01 versus pLKO.1, n = 3. B, RhoA activity in GEF-H1 knockdown (shGEF-H1) and control (pLKO.1) cells was assessed by RhoA G-LISA. *, p < 0.05 versus pLKO.1, n = 4. The blot shows the amount of total RhoA in the lysates. C, cytotoxicity was quantified by lactate dehydrogenase release (see “Experimental Procedures”). Control and knockdown cells were stimulated with increasing concentrations of complement (NS, 1, 2.5, 5, and 10%). **, p < 0.01 versus control, n = 4. a.u., arbitrary units; KD, knockdown. Error bars, S.E.