Supporting information for Biochemistry, 1995, 34(34), , DOI: /bi00034a013

Transcription

1 Supporting information for Biochemistry, 1995, 34(34), , DOI: /bi00034a013 LESNIK Terms & Conditions Electronic Supporting Information files are available without a subscription to ACS Web Editions. The American Chemical Society holds a copyright ownership interest in any copyrightable Supporting Information. Files available from the ACS website may be downloaded for personal use only. Users are not otherwise permitted to reproduce, republish, redistribute, or sell any Supporting Information from the ACS website, either in whole or in part, in either machine-readable form or any other form without permission from the American Chemical Society. For permission to reproduce, republish and redistribute this material, requesters must process their own requests via the RightsLink permission system. Information about how to use the RightsLink permission system can be found at Copyright 1995 American Chemical Society

2 K- I08IS AdPy(DR-RD) Effect of dpy fraction on relative stability of hybrid pairs. Relationship between the difference in T M values for matched pairs of DR and RD hybrids and the difference in dpy fraction in their DNA strands. AT M values for Groups A, B, C ( ) and Group D (O). Solid line are linear fits of the data for Groups A, B, and C. In this and subsequent figures, error bars are plotted only if they were greater than the size of the symbols. Numbers near the markers designate numbers of the sequences in Table I. Datafrom (Hall & McLaughlin, 1991) (T) and from (Ratmeyer et al., 1994) ( ), andfrom (Hung et al, 1994) (A).

3 Kr \Üß& /TloL Effect of deoxypyrimidine content in hybrid on relative hybrid stability compared to RNA (a) and DNA (b) counterparts. AT M values for Groups A, B, C ( ) and for Group D (O). Solid lines are drawn through the data for Groups A, B, and C. Numbers near the markers designate numbers of the sequences in Table I. Datafrom (Ratmeyer et al. 1994) ( ) andfrom (Hall & McLaughlin, 1991) (T).

4 o E > 4» Q) /N X 10 2 (number of b.p.) 1 Relationship between length and relative mobility of DNA (A) and RNA ( ) duplexes on native 20% Polyacrylamide gel. Numbers near the markers designate the numbers of sequences in Table I.

5 IMOglS 2.00 Q. o EE «u (5 < Q. JO O S 1.20 CO E J* o <5 < A.T/U b.p. content in duplex Effect of A-T/U content on free energy of RNA ( ), DNA (A), DR (0), and RD (O) duplex formation. Free energies for duplexes of Group B (a) and Group C (b). Numbers near the markers designate the numbers of sequences in Table I. (Hybrids DR ( ) and RD ( ) of seq. 14).

6 Table 2. E-selectin antisense oligonucleotides* (Bennet et al., 1994) ISIS Position Oligonucleotide sequence % Control % dpy % Number expression A+T '-UTR GAA GTC AGC CAA GAA CAG CT 50.2 ± '-UTR TAT AGG AGT TTT GAT GTG AA 73.8 ± '-UTR CTG CTG CCT CTG TCT CAG GT 6.6 ± '-UTR ACA GGA TCT CTC AGG TGG GT 20.2 ± AUG Codon AAT CAT GAC TTC AAG AGT TCT 53.4± AUG Codon TGA AGC AAT CAT GAC TTC AAG Intron/Exon: coding CCA AAG TGA GAG CTG AGA GA Coding CTG ATT CAA GGC TTT GGC AG Intron/Exon: 3'-UTR TTC CCC AGA TGC ACC TGT TT MJTR GGG CCA GAG ACC CGA GGA GA '-UTR CAC AAT CCT TAA GAA CTC TTT '-UTR GTA TGG AAG ATT ATA ATA TAT MJTR GAC AAT ATA CAA ACC TTC CAT MJTR ACG TTT GGC CTC ATG GAA GT '-UTR GGA ATG CAA AGC ACA TCC AT '-UTR TCC TCT CTT CCA GAG CAC CC '-UTR ACC TCT GCT GTT CTG ATC CT '-UTR ACC ACA CTG GTA TTT CAC AC * Oligonucleotides designed to hybridize to different regions of E-selectin mrna were synthesized containing a phosphorothioate backbone. Table 3. VCAM-1 Antisense oligonucleotides* ISIS Position Oligonucleotide sequence % Control % % Number expression dpy A+T '-UTR CGA TGC AGA TAC CGC GGA GT 37.2 ± '-UTR GCC TGG GAG GGT ATT CAG CT 58.0± '-UTR CCT GTG TGT GCC TGG GAG GG 88.4 ± AUG codon GGC ATT TTA AGT TGC TGT CG 33.6 ± Coding CAG CCT GCC TTA CTG TGG GC 66.7 ± Coding CTT GAA CAA TTA ATT CCA CCT 37.1 ± Coding/Intron TTA CCA TTG ACA TAA AGT GTT 52.4 ± Coding CTG TGT CTC CTG TCT CCG CT 26.6 ± Coding GTC TTT GTT GTT TTC TCT TCC 41.5 ± Termination TGA ACA TAT CAA GCA TTA GC 52.6 ± '-UTR GCA ATC TTG CTA TGG CAT AA 47.7 ± '-UTR CCC GGC ATC TTT ACA AAA CC 44.9 ± '-UTR AAC CCA GTG CTC CCT TTG CT 21.3 ± '-UTR AAC ATC TCC GTA CCA TGC CA 24.6 ± '-UTR GGC CAC ATT GGG AAA GTT GC 29.3 ± * Oligonucleotides designed to hybridize to different regions of VCAM-1 mrna were synthesized containing a phosphorothioate backbone.