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Inhibition of DDAH1, but not DDAH2, results in apoptosis of a human trophoblast cell line in response to TRAIL.

Lumicisi BA, Cartwright JE, Leslie K, Wallace AE, Whitley GS - Hum. Reprod. (2015)

Bottom Line: Inhibiting the expression of DDAH1, but not DDAH2, resulted in a significant increase in the sensitivity of the EVT cell line SGHPL-4 to tumour necrosis factor related apoptosis inducing ligand (TRAIL) induced apoptosis (P < 0.01).A.E.W. was supported by a grant from the Wellcome Trust (091550).There are no competing interests and the authors have no conflict interest to declare.

View Article: PubMed Central - PubMed

Affiliation: Reproductive and Cardiovascular Research Group, Institute of Cardiovascular and Cell Sciences, St George's University of London, Cranmer Terrace, London SW17 0RE, UK.

No MeSH data available.


Related in: MedlinePlus

The effect of inhibiting the expression of DDAH1 and 2 on TRAIL-induced apoptosis. Time lapse apoptosis analysis was carried out on cells transfected with either non-targeting siRNA or siRNA targeted to DDAH1 or 2, following addition of 500 ng/ml TRAIL for 24 h. Twenty cells were chosen at random for each condition and induction of apoptosis was followed by characteristic morphological changes (A). The image on the left shows a cell prior to the onset of apoptosis, the middle image shows the same cell rounding up and becoming bright before membrane blebs and blisters develop. The image on the right shows the formation of apoptotic bodies. Kinetics curves were obtained following the induction of apoptosis and the areas under the curve (AUC) were calculated (B). DDAH1 knockdown resulted in a significantly increased susceptibility to TRAIL-induced apoptosis. The data are shown as mean + SEM, from at least three independent experiments, *P < 0.05, **P < 0.01.
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DEV138F2: The effect of inhibiting the expression of DDAH1 and 2 on TRAIL-induced apoptosis. Time lapse apoptosis analysis was carried out on cells transfected with either non-targeting siRNA or siRNA targeted to DDAH1 or 2, following addition of 500 ng/ml TRAIL for 24 h. Twenty cells were chosen at random for each condition and induction of apoptosis was followed by characteristic morphological changes (A). The image on the left shows a cell prior to the onset of apoptosis, the middle image shows the same cell rounding up and becoming bright before membrane blebs and blisters develop. The image on the right shows the formation of apoptotic bodies. Kinetics curves were obtained following the induction of apoptosis and the areas under the curve (AUC) were calculated (B). DDAH1 knockdown resulted in a significantly increased susceptibility to TRAIL-induced apoptosis. The data are shown as mean + SEM, from at least three independent experiments, *P < 0.05, **P < 0.01.

Mentions: To determine whether inhibition of DDAH expression had any effect on TRAIL-induced SGHPL-4 cell apoptosis, cells were transfected with control siRNA, or siRNA to either DDAH1 or 2. Cells were stimulated with 500 ng/ml TRAIL and the induction of apoptosis was followed by time-lapse microscopy over 24 h. There was no significant change in the sensitivity of SGHPL-4 cells in which the expression of DDAH2 mRNA was inhibited. A small (but not significant) increase in apoptosis was seen with DDAH1 inhibition even in the absence of an exogenous apoptotic stimulus. However, a significant increase in apoptosis was seen after treatment with TRAIL in the cells with inhibited DDAH1 compared with those with the siRNA control (Fig. 2).Figure 2


Inhibition of DDAH1, but not DDAH2, results in apoptosis of a human trophoblast cell line in response to TRAIL.

Lumicisi BA, Cartwright JE, Leslie K, Wallace AE, Whitley GS - Hum. Reprod. (2015)

The effect of inhibiting the expression of DDAH1 and 2 on TRAIL-induced apoptosis. Time lapse apoptosis analysis was carried out on cells transfected with either non-targeting siRNA or siRNA targeted to DDAH1 or 2, following addition of 500 ng/ml TRAIL for 24 h. Twenty cells were chosen at random for each condition and induction of apoptosis was followed by characteristic morphological changes (A). The image on the left shows a cell prior to the onset of apoptosis, the middle image shows the same cell rounding up and becoming bright before membrane blebs and blisters develop. The image on the right shows the formation of apoptotic bodies. Kinetics curves were obtained following the induction of apoptosis and the areas under the curve (AUC) were calculated (B). DDAH1 knockdown resulted in a significantly increased susceptibility to TRAIL-induced apoptosis. The data are shown as mean + SEM, from at least three independent experiments, *P < 0.05, **P < 0.01.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4507332&req=5

DEV138F2: The effect of inhibiting the expression of DDAH1 and 2 on TRAIL-induced apoptosis. Time lapse apoptosis analysis was carried out on cells transfected with either non-targeting siRNA or siRNA targeted to DDAH1 or 2, following addition of 500 ng/ml TRAIL for 24 h. Twenty cells were chosen at random for each condition and induction of apoptosis was followed by characteristic morphological changes (A). The image on the left shows a cell prior to the onset of apoptosis, the middle image shows the same cell rounding up and becoming bright before membrane blebs and blisters develop. The image on the right shows the formation of apoptotic bodies. Kinetics curves were obtained following the induction of apoptosis and the areas under the curve (AUC) were calculated (B). DDAH1 knockdown resulted in a significantly increased susceptibility to TRAIL-induced apoptosis. The data are shown as mean + SEM, from at least three independent experiments, *P < 0.05, **P < 0.01.
Mentions: To determine whether inhibition of DDAH expression had any effect on TRAIL-induced SGHPL-4 cell apoptosis, cells were transfected with control siRNA, or siRNA to either DDAH1 or 2. Cells were stimulated with 500 ng/ml TRAIL and the induction of apoptosis was followed by time-lapse microscopy over 24 h. There was no significant change in the sensitivity of SGHPL-4 cells in which the expression of DDAH2 mRNA was inhibited. A small (but not significant) increase in apoptosis was seen with DDAH1 inhibition even in the absence of an exogenous apoptotic stimulus. However, a significant increase in apoptosis was seen after treatment with TRAIL in the cells with inhibited DDAH1 compared with those with the siRNA control (Fig. 2).Figure 2

Bottom Line: Inhibiting the expression of DDAH1, but not DDAH2, resulted in a significant increase in the sensitivity of the EVT cell line SGHPL-4 to tumour necrosis factor related apoptosis inducing ligand (TRAIL) induced apoptosis (P < 0.01).A.E.W. was supported by a grant from the Wellcome Trust (091550).There are no competing interests and the authors have no conflict interest to declare.

View Article: PubMed Central - PubMed

Affiliation: Reproductive and Cardiovascular Research Group, Institute of Cardiovascular and Cell Sciences, St George's University of London, Cranmer Terrace, London SW17 0RE, UK.

No MeSH data available.


Related in: MedlinePlus