Anti zcchc11

Immunohistochemistry analysis was performed applying TMA comprising human patient derived liver cancer tissue samples as described before [70 (link),74 (link),75 (link)]. Immunohistochemistry staining was analyzed semiquantitatively and the according scores were established for each antibody as described [17] (link).
Immunofluorescence assays were performed as described [76] (link). After seeding the cells in chamber slides (20 000 cells per chamber) and 24 h incubation, cells were fixed with methanol-acetic acid and afterward permeabilized using 0.1% Triton-X-100. Subsequently, the cells were blocked for 1 h using 1% bovine serum albumin/PBS, then incubated over night with specific antibodies. The following primary antibodies were used for immunohistochemistry / immunofluorescence: anti-LIN28A (1:100 dilution) (AG Meister, Laboratory for Biochemistry, University of Regensburg, Germany [28 (link),29 (link)]; anti-ZCCHC11 (1:50 dilution) (Proteintech, Rosemont, USA). For staining of the nucleus, DAPI (1:1,000 in 1mg/mL stock solution in 3 % BSA/PBS, Sigma-Aldrich Chemie GmbH, Steinheim, Germany) was used.

Cell lysis, protein isolation and Western blots were carried out as specified elsewhere [70 (link),74 (link),75 (link)]. The following primary antibodies were used: Anti-β-actin (1:5,000 dilution) (Sigma-Aldrich, St. Louis, USA), anti-ZCCHC11 (1:1,000 dilution) (Proteintech, Rosemont, USA), anti-LIN28A (1:1,000 dilution) (AG Meister, Laboratory for Biochemistry, University of Regensburg, Germany [28 (link),29 (link)]. β-Actin served as a reference for normalization. Immunoreactions were visualized applying the BCIP/NBT kit (Invitrogen by Thermo Fisher Scientific, Waltham, MA, USA). Densitometric analysis of scanned Western blot images was performed using "ImageJ" (National Institutes of Health, Bethesda, MD, USA).