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. 2016 May 20;64(3):880–893. doi: 10.1002/hep.28599

Figure 6.

Figure 6

GSH reduction causes transient cholangiocyte spheroid disruption. (A) GSH levels were measured in cholangiocytes in 2D culture over 24 hours after treatment with BSO. Data are presented as the mean ± standard error of the mean. *P < 0.05 (versus 0 time point). Representative of three independent experiments, each of which was performed in duplicate. (B) Cholangiocyte spheroids were treated with BSO for 1‐24 hours as indicated, then stained for F‐actin (green) and β1 integrin (red). Nuclei were stained with DAPI (blue). Cellular tubulin changes, F‐actin redistribution, monolayer disruption, and loss of lumens followed the same sequence and time course as for biliatresone. Scale bars: 10 μm. (C) Spheroids were treated with vehicle (DMSO) or BSO for 24 hours (two left bars) or with BSO for 1 hour followed by a 12‐ or 24‐hour washout period (two right bars). Lumens were scored as open, closed, or partially open as described in Materials and Methods (n = 286‐327 spheroids per condition).