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. 2013 Mar 11;8(3):e58871. doi: 10.1371/journal.pone.0058871

Figure 2. Effect of steviol and its derivatives on cyst progression in MDCK cyst model.

Figure 2

(A) Inhibitory effect of steviol and its derivatives on MDCK cyst formation. MDCK cysts were incubated with 100 µM of steviol and its derivatives in media containing forskolin (10 µM) after cell seeding on day 0 onward. The graph represents percent of cyst colonies at day 6 after MDCK cell seeding in the absence (control) and presence of all compounds (mean±SE; n = 4 wells/condition; **P<0.01 compared with control). (B) Inhibitory effect of steviol and its derivatives on MDCK cyst growth. The graph shows the outer cyst diameter at day 12 (mean±SE; n = 32−77 cysts; **P<0.01 compared with control). (C) Dose-response of effect of steviol on MDCK cyst growth. After cell seeding in 3D collagen gel for 6 days, media containing forskolin and steviol at doses of 50, 100, and 200 µM were added to the MDCK cells from day 6 onward. Results were shown as mean value of cyst diameter at days 6, 8, 10, and 12 (n = 43−77 cysts; **P<0.01 compared with control). (D) Representative light micrographs show MDCK cyst growth in 3D collagen gel after seeding of MDCK cells for 6 days. Three independent experiments were performed. Forskolin (10 µM) without (2D, top) or with steviol (100 µM) (2D, middle, bottom) was added to the culture medium at day 6. To test for reversibility, steviol was removed at day 9 (2D, bottom). Scale bar = 100 µm; magnification = ×10.