HGAL augments transcriptional effects and transformation potential of RhoA. (A) Raji lymphoma cells were transfected with SRF-driven luciferase reporter construct pSRE-Luc and pRL-TK plasmids and with either siRNA for HGAL or scrambled control siRNA. Forty-eight hours after transfection the cells were starved for 8 hours and then either plated on FN-coated or noncoated plates for 60 minutes followed by determination of luciferase activity. Numbers refer to luciferase activities representing means + SD of the mean of 3 independent experiments, each performed in triplicate. * indicate statistically significant (all below P < .01) difference. (B) HeLa cells stably transfected with pcDNA3.1-HGAL or pcDNA3.1-mock plasmids were transiently transfected with pSRE-Luc and pRL-TK. Forty-eight hours after transfection the cells were starved for 8 hours and then either plated on FN-coated or noncoated plates for 60 minutes and luciferase activity was determined as described in panel A. (C-D) NIH 3T3 cells were transfected with pcDNA3.1-mock, or pcDNA3.1-RhoAQ63L, pcDNA3.1-HGAL and pcDNA3.1-PDZ-RhoGEF alone or together with pcDNA3.1-HGAL (C) or pcDNA3.1-HGAL with control or RhoA siRNA (D). Cells were stained 3 weeks after transfection. Representative plates are depicted. Foci of transformation were counted in triplicate plates in independent transfections and the number of foci per μg of transfected DNA for each condition is shown as mean ± SD of the mean. * indicate statistically significant difference (all below P < .001 in panel C and P = .016 in panel D).