The entire experiment was repeated three independent times. Total protein was isolated from frozen thawed RL95 2 cells using complete RIPA buffer. meanwhile Isolated protein was resolved onto an Inhibitors,Modulators,Libraries SDS PAGE gel and transferred to a PVDF membrane. The WesternBreeze Chromogenic kit was utilized for immunodection as per the manufacturers instructions. Primary antibody concentration for western immunoblotting were the following BCL2 0. 004 ug uL. BAX 0. 004 ug uL. p BAD 0. 008 ug uL. and BAD 0. 001 ug uL. BCL2, BAD and serine 136 phosphorylated BAD primary antibodies were obtained from Santa Cruz Biotechnolgy, Inc. BAX primary antibody was obtained from Sigma Aldrich, Inc. Statistical analysis The Shapiro Wilk test was utilized to test the data for normal distribution.

All data were normally distributed except for cell proliferation and JC 1 data. Normally distributed data were analyzed using one way or two way ANOVA, when appropriate, followed by Fishers LSD test for pairwise comparison. Data that tested to be non parametric were analyzed by Friedmans one way or two way non parametric ANOVA, Inhibitors,Modulators,Libraries when appropriate, followed by Tukeys HSD test for pairwise comparison. The threshold of significance was fixed at P 0. 05. Data are presented as least square means Inhibitors,Modulators,Libraries standard error of the mean. Results Effect of L arginine on endometrial RL95 2 cell proliferation The presence of L arginine at physiological and supraphysiological concentrations increased endometrial RL95 2 cell proliferation at days 2 and 4 post treatment with proliferation being increased by approximately 4 fold on day 4.

Additionally, a dose dependent effect of L arginine Inhibitors,Modulators,Libraries on endometrial RL95 2 cell proliferation was observed on day 2 post treatment Inhibitors,Modulators,Libraries at which time cell proliferation was greater for cells treated with 800 umol L L arginine compared to those exposed to 200 umol L. Inhibitory Effect of nor NOHA on endometrial RL95 2 cell proliferation To test whether polyamines, L arginine metabolites, are responsible for L arginines effect on cell proliferation, cells were exposed to L arginine and the arginase inhibi tor nor NOHA. As in experiment one, the addition of L arginine increased endometrial RL95 2 cell proliferation, but this effect was reduced 2 fold with the addition of 800 umol L nor NOHA. Inhibitory effect of 7 NI on endometrial RL95 2 cell proliferation Cells were exposed to L arginine and the NOS inhibitor 7 NI to determine if L arginine enhances endometrial RL95 2 cell proliferation http://www.selleckchem.com/products/Oligomycin-A.html through NO biosynthesis. Again, L arginine increased endometrial RL95 2 cell proliferation, and this effect on cell proliferation was reduced with the addition of 100 umol L of 7 NI.