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. Author manuscript; available in PMC: 2014 Jul 3.

Published in final edited form as: Methods Mol Biol. 2010;602:105–117. doi: 10.1007/978-1-60761-058-8_7

Fig. 1A and Fig 1B — Flow cytometric analysis of MHC class I and II expression. Panel A. Analysis of MHC class I (H-2 K^d) and MHC class II (I-A^g7) in spleen and bone marrow cells. While nearly all NOD +/+ splenocytes and bone marrow cells express MHC class I (H-2 K^d), <1% of NOD-scid IL2rγ^null β2m^null bone marrow cells or splenocytes are MHC class I positive. Approximately 50% of NOD +/+ splenocytes and 13% of NOD +/+ bone marrow cells express MHC class II. NOD-scid IL2rγ^null as well as NOD-scid IL2rγ^null β2m^null mice have reduced percentages of MHC class II positive cells, consistent with a lack of mature B cells. Panel B. Analysis of MHC class I (H-2 K^d, NOD-derived and H-2D^b, C57BL/6-derived) and MHC class II (IA^g7, NOD-derived and I-A^b, C57BL/6-derived) expression in splenic cells. Different sets of antibodies were used because genetic backcrossing of the MHC class II targeted mutation in 129 ES cells crossed onto the C57BL/6 J strain and subsequently crossed onto the NOD strain results in the crossing of the entire MHC region onto NOD. Thus, NOD MHC class II (I-A)^null mice express the 129/C57BL/6 class I haplotype (H-2^b) and fail to express either the NOD MHC class I (H-2 K^d) or the NOD MHC class II (I-A^g7) antigens. NOD +/+ mice express H-2 K^d and H-2D^b MHC class I alleles. They also express I-A^g7 and do not express the IA^b.allele. NOD-scid IL2rγ^null mice express the H-2 K^d and H-2D^b MHC class I alleles. Expression is duller because of the absence of MHC class I-bright lymphocytes. These mice have low numbers of I-A^g7 expressing cells and lack expression of I-A^b on their cells. NOD-scid IL2rγ^nullAb^null mice lack MHC class II expression and exhibit dull H-2D^b expression with no expression of H-2 K^d.

Fig. 1A and Fig 1B — Flow cytometric analysis of MHC class I and II expression. Panel A. Analysis of MHC class I (H-2 K^d) and MHC class II (I-A^g7) in spleen and bone marrow cells. While nearly all NOD +/+ splenocytes and bone marrow cells express MHC class I (H-2 K^d), <1% of NOD-scid IL2rγ^null β2m^null bone marrow cells or splenocytes are MHC class I positive. Approximately 50% of NOD +/+ splenocytes and 13% of NOD +/+ bone marrow cells express MHC class II. NOD-scid IL2rγ^null as well as NOD-scid IL2rγ^null β2m^null mice have reduced percentages of MHC class II positive cells, consistent with a lack of mature B cells. Panel B. Analysis of MHC class I (H-2 K^d, NOD-derived and H-2D^b, C57BL/6-derived) and MHC class II (IA^g7, NOD-derived and I-A^b, C57BL/6-derived) expression in splenic cells. Different sets of antibodies were used because genetic backcrossing of the MHC class II targeted mutation in 129 ES cells crossed onto the C57BL/6 J strain and subsequently crossed onto the NOD strain results in the crossing of the entire MHC region onto NOD. Thus, NOD MHC class II (I-A)^null mice express the 129/C57BL/6 class I haplotype (H-2^b) and fail to express either the NOD MHC class I (H-2 K^d) or the NOD MHC class II (I-A^g7) antigens. NOD +/+ mice express H-2 K^d and H-2D^b MHC class I alleles. They also express I-A^g7 and do not express the IA^b.allele. NOD-scid IL2rγ^null mice express the H-2 K^d and H-2D^b MHC class I alleles. Expression is duller because of the absence of MHC class I-bright lymphocytes. These mice have low numbers of I-A^g7 expressing cells and lack expression of I-A^b on their cells. NOD-scid IL2rγ^nullAb^null mice lack MHC class II expression and exhibit dull H-2D^b expression with no expression of H-2 K^d.