Fig 4.

Effect of NPY on hESC proliferation. H9 hESCs were cultured for 4 days under feeder-free conditions using CM or UM containing 0.5 μM NPY, 0.5 μM NPY agonists, 3 μM NPY antagonists (BIBP3226 or L152804), 10 μM kinase inhibitors (AKT inhibitor, ERK1/2 inhibitor (U0126), or PKA inhibitor (H89)) in comparison with drug vehicle control (0.1% DMSO) or no treatment as indicated. Peptides and antagonists were added to fresh medium daily. The proliferation rate of H9 hESCs cultured using the indicated media was measured using BrdU incorporation. Left panels: representative images of BrdU⁺ cells. Bar = 100 mm. Right panels: quantification of BrdU⁺ cells. The relative number of BrdU⁺ cells per field of vision was quantified and is presented as the percentage of the total number of cells counted. The data are presented as the mean ± S.E. (n= 3). *P < 0.01, by t-test.