Immunohistochemistry and immunohistofluorescence in paraplast sections and cryosections, and whole-mount immunohistofluorescence were performed as previously described [40 (link),49 (link)]. For anti-PHB1 and anti-H3S10-P, a step of heat induced epitope retrieval was included after re-hydration, by boiling the slides for 20 minutes in a microwave in a solution of 10 mM sodium citrate, pH 6.0 with 0.1% Triton X-100.
Primary antibodies used were Anti-PHB1 (rabbit polyclonal, Sigma-Aldrich HPA003280, 1:100 dilution), anti-phospho-histone H3 (Ser10) (rabbit polyclonal, Cell Signaling Technology, Frankfurt/Main, Germany, code 9701, 1:100 dilution), anti-FMRFamide (Immunostar, Hudson, USA, code 20091), anti-HMW-tropomyosin ([49 (link),50 (link)], 1:500 dilution) and anti-acetylated tubulin (mouse monoclonal, clone 6-11B-1, Santa Cruz Biotechnology, Heidelberg, Germany, 1:100 dilution). In the case of anti-PHB1, we also performed a Western blot analysis with protein extracts of E. multilocularis metacestodes that confirmed that the antibody recognized a protein of the expected size. Secondary antibodies used were anti-mouse conjugated to FITC, anti-rabbit conjugated to FITC and anti-rabbit conjugated to peroxidase (Jackson ImmunoResearch, West Grove, PA, USA).
Primary antibodies used were Anti-PHB1 (rabbit polyclonal, Sigma-Aldrich HPA003280, 1:100 dilution), anti-phospho-histone H3 (Ser10) (rabbit polyclonal, Cell Signaling Technology, Frankfurt/Main, Germany, code 9701, 1:100 dilution), anti-FMRFamide (Immunostar, Hudson, USA, code 20091), anti-HMW-tropomyosin ([49 (link),50 (link)], 1:500 dilution) and anti-acetylated tubulin (mouse monoclonal, clone 6-11B-1, Santa Cruz Biotechnology, Heidelberg, Germany, 1:100 dilution). In the case of anti-PHB1, we also performed a Western blot analysis with protein extracts of E. multilocularis metacestodes that confirmed that the antibody recognized a protein of the expected size. Secondary antibodies used were anti-mouse conjugated to FITC, anti-rabbit conjugated to FITC and anti-rabbit conjugated to peroxidase (Jackson ImmunoResearch, West Grove, PA, USA).
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