American Society of Hematology

Figure 2

$Figure 2. PGE2 promotes the tube formation by activating EP4. (A) Effect of EP agonists on the tube formation. HMVECs were stimulated with PGE2, sulprostone (Sulp), butaprost (Buta), or PGE1-OH (PGE1). Tubular networks were quantified with ImageJ software (National Institutes of Health). Each point represents the mean ± SEM of values obtained from 3 independent experiments performed in triplicate. (B) Effect of EP antagonists on the tube formation. HMVECs were treated with AH23848 or AH6809 in the presence or absence of PGE2 or PGE1-OH. Tubular networks were quantified with ImageJ software (National Institutes of Health). Each point represents the mean ± SEM of values obtained from 3 independent experiments performed in triplicate. (C) Knockdown of endogenous EP4 expression. HMVECs were transfected with control scrambled siRNA (siCon) or siRNA targeting EP4 (siEP4) and allowed to grow for 60 hours. Total RNA was extracted and subjected to quantitative real-time PCR to determine expression of the EP4 gene. Expression of EP4 gene in the siEP4 sample is presented as a fraction of that in the siCon sample. Each point represents the mean ± SEM of 3 independent experiments. (D) Knockdown of EP4 expression inhibits the PGE2-induced tube formation. Representative images were obtained after 78 hours of transfection with siRNA. (E) Quantification of the tube formation after EP4 knockdown. Each point represents the mean ± SEM of triplicate samples, and experiments were repeated 3 times. ##P < .01 versus corresponding treatment in the siCon group. (A-C,E) *P < .05 and **P < .01, in comparison to corresponding control groups.$

PGE2 promotes the tube formation by activating EP4. (A) Effect of EP agonists on the tube formation. HMVECs were stimulated with PGE2, sulprostone (Sulp), butaprost (Buta), or PGE1-OH (PGE1). Tubular networks were quantified with ImageJ software (National Institutes of Health). Each point represents the mean ± SEM of values obtained from 3 independent experiments performed in triplicate. (B) Effect of EP antagonists on the tube formation. HMVECs were treated with AH23848 or AH6809 in the presence or absence of PGE2 or PGE1-OH. Tubular networks were quantified with ImageJ software (National Institutes of Health). Each point represents the mean ± SEM of values obtained from 3 independent experiments performed in triplicate. (C) Knockdown of endogenous EP4 expression. HMVECs were transfected with control scrambled siRNA (siCon) or siRNA targeting EP4 (siEP4) and allowed to grow for 60 hours. Total RNA was extracted and subjected to quantitative real-time PCR to determine expression of the EP4 gene. Expression of EP4 gene in the siEP4 sample is presented as a fraction of that in the siCon sample. Each point represents the mean ± SEM of 3 independent experiments. (D) Knockdown of EP4 expression inhibits the PGE2-induced tube formation. Representative images were obtained after 78 hours of transfection with siRNA. (E) Quantification of the tube formation after EP4 knockdown. Each point represents the mean ± SEM of triplicate samples, and experiments were repeated 3 times. ##P < .01 versus corresponding treatment in the siCon group. (A-C,E) *P < .05 and **P < .01, in comparison to corresponding control groups.

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