XB-IMG-151839
Xenbase Image ID: 151839
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Fig. S1. Structure, sequence, subcellular localization and histone methyltransferase (HMTase) activity of Prdm12. (A) Schematic representation of the Xenopus laevis Prdm12 protein. (8) Amino acid sequence alignment using Clustal Omega of the predicted Xenopus laevis Prdm12 sequence with the corresponding human, mouse, chick, zebrafish and amphioxus sequences. Conserved amino acids are highlighted in blue. The overall identity between Xenopus Prdm12 and Prdm12 of the other vertebrate species is 64%-91%. The conserved PR domain (showing 92-95% identity) and the three zinc fingers (showing 99-100% identity) are indicated. The overall sequence identity of amphioxus Prdm12 with Xenopus Prdm12 is 50%. (C) Subcellular localization of a Flag-tagged Xenopus Prdm12 protein in animal pole cells of early gastrula embryos. Note the spotted staining suggesting that the protein is localized in the cell nucleus. (D) Prdm12 has HMTase activity. Core histones were used as substrate in a HMTase assay with immunopurified mouse Flag-Prdm12 protein produced in 293T cells. Mouse Flag-Prdm3 was used as a positive control. Cells transfected with empty vector were used to control for non-specific background. The results represent the average of triplicate samples plus standard deviation (SO). P < 0.05. The amount of Prdm3 and Prdm12 proteins in the extract of transfected cells (input) and in the immunoprecipitated proteins (Ip) used in the assay is shown on the western blots (arrows). Note that Prdm12 is expressed at a much lower level than Prdm3. On the autoradiograph, multiple bands are visib le for Prdm3, which likely correspond to Prdm3 degradation products. A non-specific IgG band is indicated by an arrowhead. Image published in: Thélie A et al. (2015) Copyright © 2015. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License.
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