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Additional Supporting Information may be found in the online version of this article.

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STEM_1013_sm_SuppInfo.pdf8KSupplementary Data
STEM_1013_sm_SuppFig1.pdf605KSupplemental Figure 1: L1TD1 is highly expressed in hESCs and required for hESC selfrenewal, as well as cancer cell proliferation. A) The mean expression profile of L1TD1 in Stem Cell Matrix data (Muller FJ) in parallel with known stem cell core factors OCT-4, NANOG, and SOX-2. B) Western blot validation of L1TD1 shRNA sequences using L1TD1-V5 fusionprotein overexpression in 2102Ep EC cell line and antibody against V5. C) Validation of custom made rabbit antibody 3048 in 293T cells. L1TD1-V5 fusionprotein was overexpressed in 293T cells where overexpression produces a specific band which is responding to shRNA treatment. D) Real-time PCR analysis of L1TD1 and known stem cell markers in retinoic acid induced early differentiation serie (hESC line H9). Data from two biological replicates. E) Ingenuity pathway analysis of genome-wide gene expression data indicating the cellular processes that were influenced most in response to stable L1TD1 silencing after 6 days in EC cell line NT2D1. F) Non-Radioactive Cell Proliferation assay performed with siL1TD1 and non-targeting siRNA control 1-8 days after silencing in seminoma cell line (TCam2).
STEM_1013_sm_SuppFig2.pdf128KSupplemental Figure 2: Flow cytometry analysis of pluripotency (SSEA-3, TRA-1-60) and differentiation (A2B5 and SSEA-1) markers one and three days after knockdown of OCT4 in relation to the non-targeting control (hESC line H9). Average data from two replicate cultures.
STEM_1013_sm_SuppFig3.pdf288KSupplemental Figure 3. Alignment of the putative RNA-binding region of human L1TD1 with human pORF1. RNA-recognition-motif (RRM), C-terminal domain (CTD). Conserved residues boxed in red. Residues forming the conserved salt bridges are shaded in blue. Residues providing aromatic, RNA- binding side-chains in canonical RRMs are shaded in yellow. Asterisk (*) mark residues mutated that have strong or moderate effect on RNA-binding. Data for pORF1 [1].
STEM_1013_sm_SuppFig4.pdf383KSupplemental Figure 4. Validation of L1TD1 location. A) Endogenous L1TD1 (green) stained in hESCs with polyclonal antiserum raised against total protein antigen. B) Preserum (taken before immunization) staining (negative control). C) Anti-V5 staining of overexpressed V5-tagged L1TD1 (red) in teratocarcinoma cells (2102Ep). D) EGFP-tagged overexpression of L1TD1 in hESCs (line HS401). E) L1TD1 (red) in hESCs co-stained with pluripotency surface marker TRA-1-60 (green) co-cultured with human foreskin fibroblast. DAPI staining (blue) used to visualize nucleus in all stainings. F) L1TD1 (red) double staining with P-body marker GW182 (green) in TCam2 cells G) L1TD1 (green) double staining with P-body marker DCP1A (red) in TCam2 cells H) L1TD1 (green) double staining with SG marker TIA1 (red) in TCam2 cells I) L1TD1 (red) double staining with endosome marker EEA1 (green) in TCam2 cells. Co-Locations shown in white.
STEM_1013_sm_SuppTab1.pdf60KSupplementary Table 1
STEM_1013_sm_SuppTab2.pdf51KSupplementary Table 2

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