Anti ck7

Paraffin-embedded, formalin-fixed tissues of mice liver and spleen were immunostained for Ki67, CK7 and CK20 proteins. The signal was amplified and visualized with diaminobenzidine-chromogen, followed by counterstaining with hematoxylin. Anti-Ki67 (1:50), anti-CK7 (1:50) and anti-CK20 (1:50) were purchased from Proteintech (CK7 and CK20) and Bioworld (Ki-67).

PB samples were directly smeared onto glass slides fixed in formaldehyde solution and stained using anti-resistin Ab (Santa Cruz Biotechnology, CA, USA). For liver samples, IHC analyses of resistin, cluster of differentiation 68 [CD68, marker of Kupffer cells (KCs), i.e., liver macrophages]16 (link), CD43 (marker of lymphocytes)17 (link), α-smooth muscle actin [marker of α-SMA-positive myofibroblasts, i.e., hepatic stellate cells (HSCs)]16 (link) and cytokeratin-7 [CK-7, marker of hepatic progenitor cells (HPCs)]18 (link) were performed using anti-resistin (Santa Cruz Biotechnology), anti-CD68, anti-CD43 (Dako, Glostrup, Denmark), anti-α-SMA (Sigma, Gillingham, Dorset) and anti-CK-7 (Proteintech Group, Inc., IL, USA) Abs, respectively. Serial sections of paraffinized liver samples were used according to the manufacturers’ protocols, as the nonparenchymal cell cytoplasm is insufficient for co-staining of resistin and cell markers in the same sections. Correlations for these proteins were assessed in parallel sections. The intensity of resistin expression by IHC was measured using ImageJ software (http://imagej.nih.gov/ij/, National Institutes of Health, USA).