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. 2021 Jan 27;3(3):100250. doi: 10.1016/j.jhepr.2021.100250

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© 2021 The Authors

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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Fig. 3 — Genetic reduction of p16 reduces senescence markers in the Mdr2^-/- mouse.

(A) PCR genotyping showing the p16^-/- DNA construct. (KO) band represents the neomycin-resistance cassette on exon-1α of the Ink4a/Arf/Ink4b locus. (B) Schematic of the experimental design. P16^-/-, Mdr2^-/-, and crossbred p16^-/-xMdr2^-/- mice were harvested at 2 months of age. (C) Hepatic p21 mRNA expression, as assessed by RT-PCR, was reduced in p16^-/-xMdr2^-/- mice. (D) mRNA expression of senescence markers in isolated cholangiocytes, as assessed by RT-PCR, was reduced in p16^-/-xMdr2^-/- mice. (E) Representative images of p21 immunohistochemistry of p16^-/- (left), Mdr2^-/- (middle), and p16^-/-xMdr2^-/- mice (right) (magnification: 20×). A significant reduction in the number of p21-positive cholangiocytes was observed in p16^-/-xMdr2^-/- mice compared with the control group. (F) Quantification of p21-positive cholangiocytes. Each dot represents an image with at least 1 bile duct. Bars represent mean ± SD; n = 5–7. Scale bar: 50 μm (D). ∗p <0.05, ∗∗p <0.01, (Tukey’s multiple comparisons test). Mdr2, multidrug-resistance 2; RT, reverse transcription; TNF, tumour necrosis factor; WT, wild-type.