hAio1 is the most abundant Aiolos isoform in normal and malignant B cells. (A) Representative results of Aiolos isoform expression in 2 healthy subjects (1 and 2), 7 CLL (3-9), and 7 other B-cell lymphomas (10-16) patients of the set 1 after one-step RT-PCR amplification of PBMC RNA with the following primers: Aiolos forward 5′-GGCAGCGACATGGAAG-3′ (exon 1), Aiolos reverse 5′-TAGCTGATGGCGTTATTGATGG-3′ (exon 8). Porphobilinogen deaminase (PBGD) is used as internal control with the following primers: PBGD forward 5′-CTGGTAACGGCAATGCGGCT-3′, PBGD reverse 5′-GCAGATGGCTCCGATGGTGA-3′. The vertical line in band Aiolos is the result of a malfunction of the UV machine. (B) Ratio of absolute Aiolos isoform quantities obtained using reverse-transcribed quantitative polymerase chain reaction between primer pair/probe couple 1 (Applied Biosystems, Hs00232635_m1, exons 2-3 junction, hAio1-5 + hAio-del)(4,5,6) and primer pair/probe couple 2 (Applied Biosystems, Hs00918017_m1, exons 5-6 junction, hAio1 + hAio4) of each subgroup of patients and healthy subjects of the set 2 (B cells). Distinct PCR efficiencies between Aiolos TaqMan gene expression assays were corrected by absolute quantifications of known quantities of pcDNA3.1-hAio1 construct. Differences between groups were tested using one-way ANOVA and Tukey multiple comparison tests (Prism4.0c software; ns indicates nonsignificant, ie, P > .05).