Figure 3.

GnRH Decapeptide Secretion in Culture Medium of FGF8-Treated Cells

In the final differentiation step, secretion of GnRH decapeptide was detected in the culture medium of FGF8-treated cells, but not of the control cells.

(A) Four independent experiments using HEL11.4-derived cells (left) and three independent experiments from H9-derived cells (right) are shown. The insets in the control graphs show the same control values with smaller y-axis scale. See also Figure S3A.

(B) At high magnification the GnRH staining appeared in granular spots in both HEL11.4- and H9-derived cells, indicating vesicular packaging of the decapeptide. ROI, region of interest. Scale bars, 50 μm. See also Figure S3B.

(C) GnRH and an apoptosis marker, cleaved caspase-3, were not co-expressed, suggesting that GnRH-release was not due to apoptosis. HEL11.4-derived GnRH-expressing cells display normal nuclear morphology. Arrows indicate cleaved caspase-3 staining and nuclear morphology of apoptotic cell. Scale bars, 20 μm.