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Fig. 6 - Supplemental 1 Interaction of Piwil1 with Setdb1 and HP1α. (A) Western blot analysis of wild-type testis extracts using anti-mouse Setdb1 antibody. Arrow: Setdb1. (B) Western blot analysis of zebrafish HP1α expressed from bacterial expression vectors using anti-human HP1α antibody and anti-T7-tag antibody. Isopropyl-β-D-thiogalactopyranoside (IPTG) was added for culture of host cells (BL21) to induce expression of HP1α with T7-tag. Schema: construction of expression vector (pet21a). Arrow: HP1α fusion protein. (C) Western blot analysis of wild-type testis extracts using anti-human HP1α antibody. Arrow: HP1α. (D) Pull-down assay using Flag-tagged HP1α and Piwil1 or EGFP. Schema: conditions of coexpression of each protein in HEK-293A cells used for the pull-down assay.