Fig. 4.

ADPKD patient-derived tubules develop cysts and respond to treatment with 2DG and berberine. (A) A huADPKD microtubule stimulated with forskolin in the 3D collagen culture system displaying several cysts. The tubular lumen is lost. (B,C) Forskolin-stimulated tubules after treatment with (B) 2DG and (C) berberine. Both treatments markedly reduce cyst number and promote tubular lumen re-establishment. (D) Quantification of cyst number/mm²: no treatment (forskolin; 674.1 ± 46.1); 2DG (282.3 ± 18.5); berberine (275.3 ± 15.4). (E) A huADPKD microtubule engineered by resuspending cells in a Matrigel and collagen mixture and cultured in the 3D Matrigel system displaying several large cysts. The tubular lumen is lost. (F,G) Tubules engineered and cultured as in (E) after treatment with (F) 2DG and (G) berberine, display significantly reduced cyst numbers and a visible tubular lumen. (H) Quantification of cyst number/mm²: no treatment (control; 457.6 ± 11.8); 2DG (202.8 ± 7.0); berberine (238.7 ± 10.0). Data are expressed as means ± SEM from three independent experiments. Number of fields analyzed: (D) n = 14 for forskolin, n = 10 for 2DG, n = 12 for berberine; (H) n = 22 for control, n = 22 for 2DG, n = 21 for berberine. ^#P < 0.0001 versus forskolin, *P < 0.0001 versus control, °P < 0.05 versus berberine by one-way ANOVA with Tukey's multiple comparisons test. 2DG, 2-deoxy-d-glucose; DAPI, blue-stained nuclei. Scale bar 20 μm.