Supplemental Figures

Files in this Data Supplement:

• [Supplemental Figure 2] - "XMR detection in transfected cells using antibodies XMR^69-81, XMR^96-106, XMR^1-93 and XLR. A) XMR immunostaining of COS7 cells transfected with Xmr ORF (top row), AK015913 ORF (middle row) and non-transfected cells (bottom row). B) Western blot analysis of HEK 293 cells transfected with Xmr-egfp (lane 1), non-transfected cells (lane 2) and AK015913-egfp (lane 3). The egfp constructs were obtained by cloning the Xmr or AK015913 ORF into the pEGFP-N1 vector (Clontech) at the EcoRI and BamHI sites. The Xmr and AK015913 ORF constructs were synthesized by cloning the ORF with stop codon into the pEGFP-N1 vector at the EcoRI and NotI sites to remove the egfp ORF. Cells were transfected using LipofectAmine 2000 (Invitrogen) according to the manufacturer’s instructions."
• [Supplemental Figure 3] - "Immunoprecipitation of XMR by XMR-specific antibody XMR96-106 followed by western blot with XMR^69-81, XMR^96-106, XMR^1-93 and XLR antibodies under native (left panel) and denatured (right panel) lysis conditions. XMR is seen as a double band, probably due to protein degradation. For immunoprecipitation, 100mg of testis protein sample was lysed under native (1% Triton x-100, 300mM NaCl, 10mM Tris pH7.4, 5mM EDTA, 1mM, 0.2mM PMSF) or denatured (1%SDS, 10mM Tris pH7.4) conditions. Five hundred microliters of testis lysate was incubated with 5mg of antibody overnight at 4ºC. Complexes were precipitated with Protein A-Sepharose beads according to the manufacturer’s instructions (GE-Healthcare) and the supernatant run on a 12% SDS-PAGE gel."
• [Supplemental Figure 1] - "A MUSCLE (http://www.ebi.ac.uk/muscle/) multiple sequence alignment of Xmr (NM_009529.2), AK015913 and Xlr (NM_011725.2) cDNA sequences."