FIGURE 2.

Effects of Oxy210 on primary human HSC and drug distribution in mice. (A‐C) Primary human HSC were cultured in DMEM containing 0.1% FBS overnight and then pretreated with 5 or 10 µM Oxy210 as indicated for 2 h. The cells were then treated with TGF‐β (10 ng/ml) (filled bars) or vehicle control (empty bars). Expression of ACTA2 (A), CTGF (B) and GLI2 (C) was analysed by qPCR and normalized to GAPDH expression. (D) Primary human HSC were pretreated for 2 h with Oxy210 as indicated in DMEM containing 5% FBS. The cells were then treated with conditioned medium from Capan‐1 cells (CM) (filled bars) or vehicle control (empty bars). After 72 h, RNA was extracted and expression of GLI1 was measured by qPCR Data are presented as mean ±SD from 3 replicates per group. ##p < 0.01 vs. TGF‐β‐treated control; *p < 0.05 vs TGF‐β‐untreated control; **p < 0.01 vs TGF‐β‐untreated control. (E) Primary human HSC were plated at 10% confluence and treated with Oxy210 at the concentrations indicated for 5 days. Cells were then trypsinized and the number of cells determined. Data are reported as the mean of triplicate per group ±SD . (F) C57BL/6 mice were fed Oxy210 formulated in WD chow ad libitum for 96 h. At 96 h, terminal blood and liver samples were collected and the concentration of Oxy210 determined by LC/MS‐MS in serum and liver tissue. Data are presented as mean ± SD from 10 animals per group