The correlation of COX-2 and PTPRJ expression in aortic endothelial cells from whole-genome array expression analysis GSE39264.

<p>(A) Differential expression of PTPRJ from analysis of a MAEC whole-genome array. *P<0.05 versus untreated group. (B) Differential expression of COX-2 from analysis of a MAEC whole-genome array. *P<0.05 versus untreated group. (C) Correlation of COX-2 and PTPRJ expression from analysis of a MAEC whole-genome array. (Pearson correlation coefficient, −0.87; n = 22; P<0.01, two-tailed). Genes were selected from the microarray data using a threshold of 10% FDR (false discovery rate). P values for fold change between untreated and treated MAECs were determined by <i>t</i>-test; P values for fold change were adjusted using Benjamini–Hochberg.</p

Characterization of HUVECs by morphology and immunofluorescence.

<p>(A) Confluent cells exhibited typical cobblestone morphology; insert image at 200X magnification. (B and C) Immunofluorescence of samples labeled for VE-cadherin (B) or Von Willebrand factor (C). The immunoreactive positive cells were stained red and imaged at 400X magnification.</p

Injury-induced down-expression of PTPRJ is correlated with increased COX-2.

<p>(A–D) Representative sections of hematoxylin and eosin-stained arterial samples from control (A and B) and BI (C and D) rabbits. BI significantly induced neointimal hyperplasia. Images acquired at 200X magnification. (E) Morphometric data *P<0.05 versus uninjured group; n = 3 in each group. (F–G) PTPRJ and COX-2 expression quantified by qRT-PCR. Values were normalized to ß-actin expression. *P<0.05 versus uninjured group. (H) Correlation of COX-2 and PTPRJ expression (r = −0.74, P≤0.01).</p

PTPRJ expression is rescsued by the COX-2 selective inhibitor Celecoxib.

<p>(A) Western blot analysis of PTPRJ and COX-2 in protein extracts from HUVEC cells stably over-expressing COX-2 and treated for 24 h with 20 µM or 10 µM Celecoxib. Expression is normalized to GAPDH. Relative levels of PTPRJ were quantified using Image J. (B) ELISA quantification of PGE₂ in HUVEC supernatant; data is expressed as concentration of PGE₂. ** P<0.01 compared to the 0.1% DMSO-treated control group. (C) Relative COX-2 and PTPRJ expression in HUVEC cells treated with varying concentrations of Celecoxib. Expression is normalized to GAPDH. Relative levels of COX-2 and PTPRJ were quantified using Image J. *P<0.05.</p

PTPRJ expression is down-regulated by COX-2 over-expression.

<p>(A) Western blot analysis of PTPRJ and COX-2 in protein extracts from HUVEC cells transiently transfected for 48 h with COX-2 plasmids. Expression is normalized to GAPDH. Relative protein levels were quantified using Image J. (B) ELISA quantification of PGE₂ in the supernatant of HUVECs after transfection with COX-2 plasmids. The data is expressed as PGE₂ concentration. ** P<0.01 versus control group. (C) Correlation of COX-2 and PTPRJ expression in HUVECs after transfection with COX-2 plasmids. (Pearson correlation coefficient, −0.90, P<0.01). (D) Relative PTPRJ expression in HUVEC cells transfected with varying concentrations of COX-2 plasmids and quantified by qRT-PCR. Values were normalized to PRL22 expression. *P<0.05 versus control group.</p

Primers used for the real time analysis.

<p>Primers used for the real time analysis.</p