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. 2011 Jan 14;300(4):L596–L604. doi: 10.1152/ajplung.00029.2010

Fig. 4.

Fig. 4.

LPS/TNF treatment suppressed proliferation in bPAEC, and inhibiting l-arginine uptake attenuated the LPS/TNF-induced suppression of proliferation. A: bPAEC were treated with vehicle (control) or LPS/TNF and either no added l-leu or 10 mM l-leu for 24 h. The bPAEC were harvested, and viable cells were counted using Trypan blue exclusion. LPS/TNF treatment resulted in fewer viable bPAEC than in vehicle-treated bPAEC (control). The addition of 10 mM l-leu to vehicle-treated bPAEC had no apparent effect on viable cell numbers. However, the addition of 10 mM l-leu to the LPS/TNF-treated bPAEC resulted in significantly more viable cells than in bPAEC treated with LPS/TNF alone. Values are means ± SE; n = 5–6 in each group. *LPS/TNF different from control, P < 0.005. #LPS/TNF + l-leu different from TNF/LPS alone, P = 0.049. B: LPS/TNF treatment decreased viable cell number in part by increasing apoptosis. Cells were treated with vehicle (control), LPS/TNF, or LPS/TNF + l-leu (10 mM) for 24 h, and then protein was harvested for detection of cleaved poly(ADP-ribose) polymerase (PARP) and cleaved caspase-3 by Western blotting. A representative blot is shown in B. The first two lanes are from control with little cleaved PARP or caspase-3 expression, the middle lane is from LPS/TNF with substantial bands for both cleaved PARP and caspase-3, and the last two lanes are from LPS/TNF + l-leu-treated cells, and there is a substantial attenuation of both the cleaved PARP and cleaved caspase-3 bands. The β-actin is a protein loading control and demonstrates equal protein loading of the gels. C: the densitometry results for the cleaved caspase-3 Western blots normalized to β-actin. Values are means ± SE; n = 3–4 in each group. *LPS/TNF different from control, P < 0.001. #LPS/TNF + l-leu different from LPS/TNF alone, P < 0.005. D: the densitometry results for cleaved PARP Western blots normalized to β-actin. Values are means ± SE; n = 3–4 in each group. *LPS/TNF different from control, P < 0.05. #LPS/TNF + l-leu different from LPS/TNF alone, P < 0.05.