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. Author manuscript; available in PMC: 2017 Sep 1.
Published in final edited form as: Hepatology. 2016 May 20;64(3):880–893. doi: 10.1002/hep.28599

Figure 6. GSH reduction causes transient cholangiocyte spheroid disruption.

Figure 6

(a) GSH levels were measured in cholangiocytes in 2D culture over 24 h following treatment with BSO. Mean ± SEM, *p<0.05 compared to 0 time point. Representative of 3 independent experiments each with duplicates.

(b) Cholangiocyte spheroids were treated with BSO for 1–24 h as indicated, then stained for F-actin (green) and β1 integrin (red). DAPI nuclear stain (blue). Cellular tubulin changes, F-actin redistribution, monolayer disruption, and loss of lumens followed same sequence and time course as for biliatresone. Scale bar, 10 μm. (c) Spheroids were treated with vehicle (DMSO) or BSO for 24 h (two left bars), or with BSO for 1 h followed by a 12 or 24 h washout period (two right bars). Lumens were scored as open, closed, or partially open as described in the methods. N=286–327 spheroids per condition.