Figure 6. Pharmacologic and genetic inhibition of autophagy reduced PLD cholangiocyte proliferation, cyst growth and cholangioid formation of in 3-D cultures.

(A) Proliferation of PCK rat cholangiocytes was inhibited by Bafilomycin A₁ and HCQ (both, 100 nM) as assessed by MTS absorbance. (B) Representative light microscopy images and (C) bar graphs demonstrate inhibited expansion of dissected PCK rat cystic bile ducts treated with Bafilomycin A₁ and HCQ (both, 100 nM). (D) Representative western blots and quantitation of protein band density demonstrate reduced expression of Atg7 in ADPKD cholangiocytes stable transfected with ATG7 siRNA compared to cholangiocytes transfected with scrambled control. (E) Representative confocal and phase-contrast microscopy images of cholangioids formed by human ADPKD cholangiocytes in 3-D culture. ADPKD cholangiocytes transfected with ATG7-siRNA developed smaller cystic structures compared to cholangiocytes transfected with scrambled control siRNA. Data are presented as MEAN±SD. Abbreviation: Un-tr – untreated, BAF – Bafilomycin A1, HCQ – hydroxychloroquine.