Figure 4
Expression of RNF217 in primary human leukemia samples.
(a) Primary human bone marrow samples from leukemia patients were analyzed for their expression of RNF217 by RT-PCR. TBP expression was assayed as a control. (b) Microarray expression profiles of primary human bone marrow samples from leukemia patients19 were generated. Data for RNF217 expression was log2 transformed and evaluated (normal bone marrow (nBM; n = 10), CML; n = 10, CALM/AF10-positive AML (CALM_AF10; n = 10), AML with normal karyotype and FLT3 negative (AML_nk; n = 10), AML with MLL rearrangement (AML_MLL; n = 9), AML with inv(16)(p13q22) (CBFB-MYH11 fusion, AML_M4; n = 10), AML with t(15;17)(q21;q22) (PML-RARA fusion, AML_M3; n = 10), AML with t(8;21)(q22;q22) (AML1-ETO fusion, AML_M2; n = 10), AML with normal karyotype and FLT3-ITD (AML_FLT3; n = 10), AML with complex aberrant karyotype (AML_comp; n = 10), ALL with t(9;22)(q34;q11) (BCR-ABL fusion, ALL_Ph; n = 10), ALL with t(4;11)(q21;q23) rearrangement (MLL-AF4 fusion, ALL_MLL; n = 10), common ALL (ALL_BA; n = 10) and pro-B ALL (ALL; n = 10)). The probe set for RNF217 on the microarrays used (Affymetrix HGU133, probe set 235492_AT) detects exon 13 of human RNF217 (see Supplementary table 1). This exon is part of all splice variants shown in Supplementary table 2. (c) RNF217 expression in primary human bone marrow samples from leukemia patients was assessed by quantitative RT-PCR with β-actin as reference gene. Expression levels were calculated from cT values by the 2-ΔcT method (ALL; n = 19, AML; n = 10, CML; n = 10), Mean ± standard error of mean. ** P ≤ 0.01. The primers used for this experiment span exons 11 and 12 of the human RNF217 gene (see Supplementary table 1). It has to be noted that not all of the identified putatively protein-coding splice variants of RNF217 contain exon 12 (see Supplementary table 2), therefore it is possible that not all splice variants of this gene in the leukemic samples might have been detected.
