1. Leukemic Cellular Retinoic Acid Resistance and Missense Mutations in the PML-RAR Fusion Gene After Relapse of Acute Promyelocytic Leukemia From Treatment With All-trans Retinoic Acid and Intensive Chemotherapy
by Wei Ding, Yun-Ping Li, Lucio M. Nobile, George Grills, Ines Carrera, Elisabeth Paietta, Martin S. Tallman, Peter H. Wiernik, and Robert E. Gallagher
Blood
Volume 92(4):
August 15, 1998
©1998 by American Society of Hematology

2. Schematic structure of PML-RAR mRNA showing the major functional domains.
Schematic structure of PML-RAR mRNA showing the major functional domains. The PML portion is hatched and the RAR portion is clear. Numbers in the boxes indicate exons. Vertical arrows indicate the positions of break/fusion sites in the translocated PML gene that result in the formation of three different forms of PML-RAR mRNA: S-, V-, and L-forms. Arrowheads show the sites of PCR primers used in this study. Primers used for screening of the PML region are shown beneath the schematic bar, whereas those for the RAR region are shown above it. The asterisks indicate the positions of base changes found in the PML-RAR allele, and the coding region amino acid changes are shown in expanded form below the schematic bar. The abbreviations are as follows: Pro, proline-rich domain; Cys, cysteine/histidine clusters; -Helix, -helical coiled-coil dimerization domain; B-F, B- to F-region of RAR; ZF, zinc-finger RING motif; BB1 and BB2, two B-boxes; DBD, DNA binding domain; DD, dimerization domain; LBD, ligand binding domain; AF-2, activator function-2 domain; AF-2 AD CORE, 7 amino acid long AF-2 activation domain; 3′-UT, 3′ untranslated region.
Wei Ding et al. Blood 1998;92:
©1998 by American Society of Hematology

3. In vitro RA-induced differentiation of pretreatment and relapse APL cells from selected patients determined by the NBT test.
In vitro RA-induced differentiation of pretreatment and relapse APL cells from selected patients determined by the NBT test. (A) Case no. 12; (B) case no. 9; (C) case no. 17; (D) case no. 14. (▪) pretreatment sample; (•) relapse sample. Case no. 12 only: (▴) primary DA refractoriness; (▾) postintravenous liposomal RA.
Wei Ding et al. Blood 1998;92:
©1998 by American Society of Hematology

4. Comparison of the gel electrophoretic pattern of RT-PCR products from the pretreatment and relapse specimens of case no. 9.
Comparison of the gel electrophoretic pattern of RT-PCR products from the pretreatment and relapse specimens of case no. 9. PCR amplification used primer pair I as described in the Materials and Methods. M1, Hae III-digested ΦX 174 DNA; M2, 100-bp size standard; H, HL-60 cell RNA; N, NB4 cell RNA; P, pretreatment RNA; R, relapse RNA. PCR band 1, full-length V-form with PML breaksite at nt 1685; band 2, full-length product of comigrating atypical isoforms with breaksites at nts 1576 and 1581; band 3, same as band 1 but lacking exon 5 due to alternative splicing; band 4, same as band 2 but lacking exon 5; band 5, isoform lacking exons 5 and 6 due to alternative splicing.
Wei Ding et al. Blood 1998;92:
©1998 by American Society of Hematology

5. Automated DNA sequence analysis of PCR products from four APL cases with single nucleotide changes.
Automated DNA sequence analysis of PCR products from four APL cases with single nucleotide changes. The upper panels show bi-allelic sequencing results for the pretreatment samples, which contain normal signals as indicated by the arrows in (A), (B), and (C), whereas (D) demonstrates a heterozygous pattern. The middle panels show heterozygous bi-allelic signals for the corresponding relapse samples. The lower panels show the results of nested, PML-RAR allele-specific amplification of PCR products from the relapse cases. (A) Case no. 9, T→C (Met→Thr). (B) Case no. 17, C→T (Arg→Trp). (C) Case no. 14, C→G (Leu→Val). (D) Case no. 2, C→T in 3′-UT region (illustrated from antisense strand sequence analysis, ie, shown as G→A; in the upper panel the heterozygotic mutant A appears on the shoulder of the normal G).
Wei Ding et al. Blood 1998;92:
©1998 by American Society of Hematology