Fig. 9.

Apoptosis induced formation of receptor for advanced glycation end products (RAGE)-inositol monophosphatase 1 (IMPA1) complex and protein kinase B (Akt) activation in vitro. A: apoptosis induced in pulmonary artery vascular cells was used to prepare conditioned media. B and C: cell media collected from apoptotic but not untreated control cells induced interaction between RAGE and IMPA1 (B) and activation of Akt (C) in naïve human pulmonary artery smooth muscle cells (HPASMC). Akt phosphorylation was significantly attenuated in the presence of selective IMPA1 inhibitor (L-690,330; 500 µM) and RAGE antagonist (RAP; 50 µM), confirming an important role of both proteins in activation of Akt signaling; n = 6/group (C) and n = 5/group (B). Protein loading was normalized per total sample protein using stain-free imaging technology. Results are expressed as box and whisker plots (boxes: 25th to 75th percentile of the data; whiskers: minimum to maximum; line represents the median value). Statistical analysis was performed using unpaired t-test (B) or Newman-Keuls multiple-comparisons test (C). Apo, media collected from apoptotic cells. *P < 0.05 vs. control group; #P < 0.05 vs. apoptosis group.