![Figure 2. Correlation of expression of AID and its splice variant with identification of variant tumor-derived isotype switch transcripts and circle transcripts in unmutated and mutated VH gene t(11;14) mantle cell lymphoma. Mutational status of tumor VH genes is shown as % homology to germ line VH gene segment (> 98% homology denotes unmutated status [UM] vs mutated [MUT]). Variant tumor isotype transcripts were identified by amplification with CDR3-specific primers together with downstream isotype-specific CH primer (ND, not determined). Two AID transcripts, wild-type (wt; 656 bp) and variant (loss of exon 4, -ex4; 454 bp), were identified by RT-PCR and amplified bands of predicted size resolved on agarose gels prior to nucleotide sequence verification. AID expression in the lymphoma cell line Ramos served as a positive control. Amplification of VH genes was used to confirm integrity of RNA and loading. Comparable amounts of cDNA were used for VH gene and AID amplification. Weakly amplified bands for variant AID transcripts in 2 cases (MUT case 7 and UM case 12) were confirmed as positive by cloning and sequence analysis. In UM case 11, additional bands in AID amplification were nonspecific as shown by sequence data. In 4 of 4 normal PBMNC preparations, AID expression was not detected under the conditions used (data not shown).](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/103/7/10.1182_blood-2003-05-1632/6/zh80070459090002.jpeg?Expires=1722777073&Signature=XL~biyJGLZTF0pmsrV9BaAfluKXg7N8IttrNM2Npx~AsnAtatVzBDQBB~4pV~b9pvGka6wH15~gs0QgUm9jz~0-s8VUiNwZ4EMBV5JBeRP9UJvmQZYH6sWV7nIfgpG0fkqRfcbVHlmlAQ-UzoQzokG2tdFEWv5yIcIrYs9s8So9iSlfYxDm17fU3Fn~7gpTbE49phR7TY6nrs2lC4ZHp6Rd6m8WuPKytLNtSCZdBhalxnl~3SPxibJu4AlPdGEokc1qA--AYJVflqGHY94bJQY0yVnqfIVO2xlFLSZ7xQp2vcDdqL~panyRatG1X3NfkbZLtQxbvwhIIF0LD-~VVjQ__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Correlation of expression of AID and its splice variant with identification of variant tumor-derived isotype switch transcripts and circle transcripts in unmutated and mutated VH gene t(11;14) mantle cell lymphoma. Mutational status of tumor VH genes is shown as % homology to germ line VH gene segment (> 98% homology denotes unmutated status [UM] vs mutated [MUT]). Variant tumor isotype transcripts were identified by amplification with CDR3-specific primers together with downstream isotype-specific CH primer (ND, not determined). Two AID transcripts, wild-type (wt; 656 bp) and variant (loss of exon 4, -ex4; 454 bp), were identified by RT-PCR and amplified bands of predicted size resolved on agarose gels prior to nucleotide sequence verification. AID expression in the lymphoma cell line Ramos served as a positive control. Amplification of VH genes was used to confirm integrity of RNA and loading. Comparable amounts of cDNA were used for VH gene and AID amplification. Weakly amplified bands for variant AID transcripts in 2 cases (MUT case 7 and UM case 12) were confirmed as positive by cloning and sequence analysis. In UM case 11, additional bands in AID amplification were nonspecific as shown by sequence data. In 4 of 4 normal PBMNC preparations, AID expression was not detected under the conditions used (data not shown).
