Nuclear lamina changes in patients with NAFLD. (A) Nuclear immunofluorescence (lamin B1; green) and DAPI nuclear staining of frozen liver sections from patients with normal (top left) and fatty livers (top middle and right). Nuclei in individuals with normal livers have a round shape, whereas nuclei in NAFLD patients are irregular in shape and distorted. (Bottom) Representative liver sections from normal and NAFLD patients stained with H&E. Lipid accumulation is apparent on histological sections by presence of lipid droplets in NAFLD livers. (B) Western blot analysis of protein nuclear extracts from livers of three normal and six NAFLD patients with antibodies to LBR, LMNB1, LMNA/LMNC, and Histone H3 (loading control). Protein expression of LBR and lamina A (LMNA) is increased in most patients with NAFLD. Protein levels of lamin B1 (LMNB1) are decreased in some NAFLD patients. (C) Venn diagram showing the results of genome-wide location analysis for lamin B1 (ChIP-seq) in livers of normal and NAFLD patients, identifying 1345 domains in normal, 1457 in mild steatosis, and 1616 in severe steatosis, of which 1355 were called bound in all conditions by Epic. (D) Lamin B1 ChIP-seq signal (RPKM) calculated at LADs decreases significantly in NAFLD patients to the same extent in mild and severe steatosis. (E) PscanChIP identified highly enriched consensus sites for the forkhead motif in all patients.
