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. 2021 Feb 26;11(3):141. doi: 10.3390/metabo11030141

Figure 1.

Figure 1

Cholesterol supplementation induces acinar-to-ductal metaplasia (ADM) but does not significantly impact EGFR or sonic hedgehog (Shh) signaling. Primary wild-type pancreatic acinar cells were either embedded in a synthetic hydrogel cast (A,C,D,E) or cultured suspended in RPMI medium (B) and stimulated with recombinant EGF (100 ng/mL, unless otherwise indicated). A, Cells were treated with free cholesterol (10 μg/mL) or vehicle (ethanol). After 5 days, morphology of cellular clusters was examined. Figure shows representative images of triplicate experiments. Scale bar, 50 μm. B, Cells were lysated 20 min after EGF addition. Western blotting shows phosphorylation of EGFR and downstream effectors. Representative image of duplicate experiment. C–D, Acinar cells were independently isolated from 3 mice (#1, #2, #3) and treated with cholesterol and SHH inhibitor Cyclopamine or vehicle. Morphology was examined after 5 days (C) and number of acini/duct structures was quantified by a blinded investigator (in D). **, p < 0.01. Scale bar, 50 μm. E, qPCR analysis of the Ptch1 gene in cells treated like in A.