SUPPLEMENTARY MATERIAL Jens Mattow, Ilja Demuth, Gisela Haeselbarth, Peter R. Jungblut,

1. SUPPLEMENTARY MATERIAL Jens Mattow, Ilja Demuth, Gisela Haeselbarth, Peter R. Jungblut, Joachim Klose Selenium-binding protein 2, the major hepatic target for acetaminophen, shows sex differences in protein abundance Electrophoresis, submitted.

2. 6805, 6809, 7814*, 8810* Parents 5514 6613* 4303* 5205* 4301* 5216* Family 1 Mother: Calzinato Father: Aarhus F1(CxA) Hybrids Male 1 Female 5 Female 1 Female 2 Female 6 Female 3 Female 4 Parents Family 2 Mother: Aarhus Father: Calzinato F1(AxC) Hybrids Females vs. Males Male 1 Male 2 Male 3 Parents Family 3 Mother: Calzinato Father: Aarhus F1(CxA) Hybrids Female 1 Female 2 Female 3 Female 4 Female 5 Female 6 Male 1 Male 2 Parents Family 4 Mother: Aarhus Father: Calzinato F1(AxC) Hybrids Female 1 Female 2 Male 1 Male 2 Male 3 Male 4 Male 5

3. Figure legend Silver-stained 2-DE patterns of cytosolic liver proteins were investigated in 8 parental mice (4 females; 4 males) of 2 subspecies, namely Mus musculus musculus (inbred strain: Calcinato B) and Mus musculus domesticus (inbred strain: Aarhus), as well as in their offspring (25 F1 hybrids; 14 females; 11 males). The primary goal of this analysis was to identify strain-specific protein variants, which reveal a mode of inheritance compatible with the concept of genomic imprinting. In this context, the protein patterns of the parental mice(2 replicate gels per animal) were compared with each other to elucidate strain-as well as sex-dependent protein variations. The figure depicts sectors from the protein patterns of all 33 mice investigated. The 10 protein spots labeled showed very low intensities in female mice (left side), but high intensities in males (right side). Seven of these spots (marked by asterisks) were unambiguously identified as SBP2 by MALDI-MS. The remaining 3 protein spots (SSP5514, SSP6805, SSP6809) most likely represent additional SBP2 variants, but were not analyzed by MS, due to their absence in preparative CBB G250 stained 2-DE gels. Edman degradation revealed that SSP6613 represents an N-terminally truncated SBP2 variant, which lacks the amino acids 1-100 of the full-length protein. Similarly, protein spots SSP5216 and SSP5205 were identified as SBP2 variants lacking the amino acids 1-267 and 1-225, respectively. Edman degradation of SSP8810 did not reveal any sequence data suggesting that this protein spot represents an N-terminally blocked full-length SBP2 variant. For reasons of clarity the spot labels are abbreviated, e.g. protein spot SSP8810 is designated as 8810. Please note that only the parental mice were analyzed in detail to elucidate sex-dependent protein variations. However, reproducible sex differences in SBP2 abundance were also detected in the 25 F1 hybrids.