Mutation frequency of Igε transgenic regions
| Activation and organ, according to strain . | Sorting . | Mutation frequency, 10−4* . | Mutations/total nucleotides . | P† . | Deletions‡ . |
|---|---|---|---|---|---|
| Experiment 1—in vitro activation with anti-CD40 + IL-4 | |||||
| EPS21 | IgE+ | 3.0 | 6/19989 | NS | 0/6 |
| L12 | IgE+ | 6.1 | 20/33039 | NS¶/<.05** | 1/20 |
| R27 | IgE+ | 17†† | 11/6656 | <.001/<.05 | — |
| R27 | IgE+ | 13 | 26/19446 | <.05/<.001 | 4/26 |
| R27 | IgE+ | 4.3‡‡ | 3/6960 | NS | — |
| R27 | IgE+ | 8.0§§ | 9/11280 | NS/<.05 | — |
| R27 | IgE+ | 11‖‖ | 21/18950 | <.05/<.001 | — |
| Experiment 2—in vitro activation with anti-CD40 + IL-4 | |||||
| EPS21 | CFSEdull | 2.7 | 2/7391 | NS | 0/2 |
| L12 | CFSEdull | 7.3 | 6/8235 | NS | 0/6 |
| R7 | CFSEdull | 12 | 16/13584 | <.05/<.05 | 4/16 |
| Experiment 3—Peyer patches | |||||
| EPS21 | GC B cells | 5.7 | 7/12258 | NS | 0/7 |
| L12 | GC B cells | 10 | 12/11699 | NS | 0/12 |
| R27 | GC B cells | 47 | 48/10114 | <.001/<.001 | 2/48 |
| R7 | GC B cells | 18 | 29/16344 | <.05/<.05 | 0/28 |
| Activation and organ, according to strain . | Sorting . | Mutation frequency, 10−4* . | Mutations/total nucleotides . | P† . | Deletions‡ . |
|---|---|---|---|---|---|
| Experiment 1—in vitro activation with anti-CD40 + IL-4 | |||||
| EPS21 | IgE+ | 3.0 | 6/19989 | NS | 0/6 |
| L12 | IgE+ | 6.1 | 20/33039 | NS¶/<.05** | 1/20 |
| R27 | IgE+ | 17†† | 11/6656 | <.001/<.05 | — |
| R27 | IgE+ | 13 | 26/19446 | <.05/<.001 | 4/26 |
| R27 | IgE+ | 4.3‡‡ | 3/6960 | NS | — |
| R27 | IgE+ | 8.0§§ | 9/11280 | NS/<.05 | — |
| R27 | IgE+ | 11‖‖ | 21/18950 | <.05/<.001 | — |
| Experiment 2—in vitro activation with anti-CD40 + IL-4 | |||||
| EPS21 | CFSEdull | 2.7 | 2/7391 | NS | 0/2 |
| L12 | CFSEdull | 7.3 | 6/8235 | NS | 0/6 |
| R7 | CFSEdull | 12 | 16/13584 | <.05/<.05 | 4/16 |
| Experiment 3—Peyer patches | |||||
| EPS21 | GC B cells | 5.7 | 7/12258 | NS | 0/7 |
| L12 | GC B cells | 10 | 12/11699 | NS | 0/12 |
| R27 | GC B cells | 47 | 48/10114 | <.001/<.001 | 2/48 |
| R7 | GC B cells | 18 | 29/16344 | <.05/<.05 | 0/28 |
Purified splenic B cells were activated in vitro with anti-CD40 + IL-4 (experiments 1 and 2). In experiment 1, IgE+ cells were sorted on day 6. In experiment 2, cells were labeled with CFSE before culturing and the 10%-11% dullest cells were sorted on day 6. In experiment 3, GC B cells were sorted form Peyer patches.
NS indicates nonsignificant:—, not analyzed.
Mutation frequency of the middle transgenic Sε region, except when otherwise stated. Each specific mutation was counted only once, meaning that the frequency of mutations is underestimated. Significant values have been italicized.
Calculated by Fisher exact test.
Number of deletions/total number of mutations.
Compared with transgenic Sε sequence of R7 tail DNA with mutation frequency of 5.8 × 10−4 (8/13690).
Compared with transgenic Sε sequence from E 21 within the same experiment.
Mutation frequency of transgenic Iε region.
Mutation frequency of transgenic Cε region 5′ of tag.
Mutation frequency of transgenic 3′ Sε region.
Mutation frequency of transgenic 5′ Sε region.