Journal: Fertility and sterility

Article Title: Sildenafil stimulates human trophoblast invasion through nitric oxide and guanosine 3′,5′-cyclic monophosphate signaling

doi: 10.1016/j.fertnstert.2015.02.025

Figure Lengend Snippet: Trophoblast invasion from first-trimester chorionic villous explants. First-trimester villous explants were cultured on Matrigel-coated transwell inserts and treated with vehicle or 350 ng/mL sildenafil for 72 hours before fixation. Examples of outgrowths from villous explants cultured with vehicle (A, C, E) or 350 ng/mL sildenafil (B, E, F) are shown. Bright-field images (A, B) and paired fields labeled with 6-diamino-2-phenylindole (C, D) or anti-Ki67 (E, F) are included. Size bar in (A) applies to all image panels. Outgrowth length and cell proliferation were measured using Simple PCI image analysis software (G, H). The graph in (G) shows outgrowth lengths for triplicate experiments in which 10 villous explants were measured for each treatment. * P<.05 vs. vehicle. The graph in (H) shows the percentage of cell nuclei in the outgrowth regions that were labeled with anti-Ki67 antibody. The differences between vehicle- and sildenafil-treated explants were not significant (n = 8). Error bars represent SEM.

Article Snippet: Images were processed to determine gray level using Simple PCI (Hamamatsu) imaging software, as previously described ( 31 , 32 ).

Techniques: Cell Culture, Labeling, Software