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Blood, Vol. 109, Issue 1, 159-167, January 1, 2007
Regulation of  germline transcription and switch region mutations by IgH locus 3' enhancers in transgenic mice
Blood Laurencikiene et al.
109: 159
Supplemental materials for: Laurencikiene et al, Vol 109, Issue 1, 159-167
Files in this Data Supplement:
- Document S1. Materials and methods (PDF, 75.6 KB)
- Table S1. The percentage of IgG1- and IgE-producing cells in B cells from wild-type or transgenic mice (JPG, 53.2 KB)
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Splenic B cells were cultured with anti-CD40 plus IL-4 for 4 days. Cells were then washed, fixed, permeabilized, and stained with biotinylated anti-IgG1 (553441; BD Biosciences, Palo Alto, CA) or -IgE followed by streptavidin/FITC and analyzed by flow cytometry.
- Figure S1. Relative expression of GL ε transcripts (JPG, 75.7 KB)
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Endogenous and transgenic GL  transcripts were compared with Mb-1 RNA expression using real-time RT-PCR. In each individual group Mb-1 expression was set to 100%. Absolute expression is divided by copy number of the gene. (A) Relative expression of GL transcripts is shown as a percentage of Mb-1 expression. Mean values from the triplicates are shown and SDs are calculated. (B) The same experiment is shown as a graph.
- Figure S2. Mutation spectrum of the analyzed transgenic Sε region in anti-CD40 plus IL-4–activated B cells from ENH-right mice (JPG, 130 KB)
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The exchanged nucleotide is shown above the appropriate nucleotide in the germline sequence. Mutated nucleotides from IgE+ cells are shown in regular style, and those from CFSE-labeling experiments are shown in bold. Deletions are indicated by arrows, those with thin lines from IgE+ cells, those with thick lines from CFSE-labeled cells. The position of the arrowhead marks the end of the deletions. The double-headed arrows indicate the entire length of deletion. The single-headed arrows indicate longer deletions. For long deletions, the base of the arrow is marked with a dash or a dot so that the beginning and the end of the deletion can be identified. The arrows with a dot at the base indicate a recombination between two different S regions.
- Figure S3. Mutation spectrum of the analyzed Sε region from GC B cells sorted from Peyer patches of ENH-right mice (JPG, 128 KB)
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Explanation as in Figure S1.
- Figure S4. Pie chart depicting the proportion of sequences that carry 0, 1, 2, 3, or more mutations over the 681-bp region analyzed from the R7 founder in Table 1, Experiment 3 (JPG, 29.6 KB)
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The number of clones analyzed is shown in the middle.
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