Supplementary Information A thermodynamic overview of naturally occurring intramolecular DNA quadruplexes
Niti Kumar and Souvik Maiti*
Proteomics and Structural Biology Unit, Institute of Genomics and Integrative Biology, CSIR, Mall Road, Delhi 110 007, India. ∗To whom correspondence should be addressed. Email:
[email protected], Ph no: +9111-2766-6156. Fax: +91-11-2766-7471.
SI Table 1: Sequences having different loop length used in the study with their respective CD signature obtained in 10 mM Sodium cacodylate buffer, pH 7.4 with 100 mM KCl at 25°C.
Oligo Name
Gene
Q1 Q2 Q3 Q4 Q5 Q6 Q7 Q8 Q9 Q10 Q11 Q12 Q13 Q14 Q15 Q16 Q17 Q18 Q19 Q20 Q21 Q22 Q23 Q24 Q25 Q26 Q27 Q28 Q29 Q30 Q31 Q32 Q33 Q34 Q35 Q36 Q37 Q38 Q39 Q40 Q41
VEGF WNT 3 WNT5A PDGFB JUNB TAL1 THPO FOS PIM1 FGF 3 FES VAV1 IGF2 FOSL1 TAL1 AKT2 THRA FBXW 7 HRAS WNT1 c-MYC NFKB 2 GLI 1 HCK HCK YES 1 AKT 1 BCL 2 NOTCH 1 c-KIT JUNB SRC VAV 2 AKT 2 MYB ROS 1 TAL 1 WNT1 CTTN AXL FGF 3
Position relative to TSSa -1805 -149 -934 -714 -1695 -313 -1759 -1689 -328 -18 -431 -531 -1254 -206 -566 -79 -123 -58 -709 -1013 -913 -424 -103 -800 -795 -112 -59 -917 -168 -82 -1746 -1708 -160 -87 -797 -1323 -545 -191 -72 -55 -120
Sequence (5′-3′)
GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG
A CC A T TG G TGG GAGCT CG GGA A CA T CTG GCC GGAG CG CGGC GGGACT AAAGA GA CGC CCGGC GA CC T C CCA A TGG CT TGAGCT CGTG GGCTG CG GTGG CGG GGCCACC AGGAGGG CT GATTGAT
GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG
TTG C CA T CG CGGT AG A CG GC CT A CC CG C GC GGC GCC C CAT TGGGGA GCGC A CC CCAGAA AA CGGCTCC AGC AGCGCAA CCCCGA TT CT ACTGC GGA T AT AGC CA CA GG CTGG GAGC
GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG
T AG AA CTCT CC G A A GCG TA ACCA AACT AGCTT CGC CGCG GG GGC G CA A TG CGC CTGG CCAGAA AG ACGCCGC CGC GCGGA CGC CT CCAGGCC GCT TG GGC TGGGGCCC AGATG GGCCGC GGC C TGGAGGC AG
GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG
Total
CD signatureb
5 5 5 6 6 6 6 7 7 7 7 7 8 8 8 8 8 8 9 9 10 10 10 10 10 10 11 11 11 11 11 11 11 11 11 11 12 12 12 12 12
Parallel Parallel 1 Parallel 1 Parallel 1 Parallel 1 Parallel Parallel 1 Parallel 1 Parallel Parallel Parallel 1 Parallel 1 Parallel Parallel 1 Parallel 1 Parallel Parallel Parallel Parallel Parallel 1 Parallel 1 Parallel 1 Parallel 1 Parallel 1 Mixed Mixed Mixed Mixed Mixed Parallel 1 Mixed Mixed Mixed Parallel 1 Parallel 1 Parallel 1 Mixed Parallel 1 Mixed Parallel 1 Parallel 1
Q42 Q43 Q44 Q45 Q46 Q47 Q48 Q49 Q50 Q51 Q52 Q53 Q54 Q55 Q56 Q57 Q58 Q59 Q60 Q61 Q62
MAF MPL MYB ABL 1 MYEOV VEGF FLI 1 FGR WNT 3 CSF 1R THRA FES REL FGF 4 BCL 3 KRAS THPO RALB EGFR VAV 1 FGF 6
-446 -35 -342 -648 -742 -89 -223 -131 -32 -1105 -135 -237 -196 -267 -1872 -327 -272 -1755 -300 -501 -272
a
GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG
TGGA CTGGCCT AGGCGGC CC TAAA AG AGGAAGA CTTTT CT C CA GGA CGCGGCA CCCTG ACAGGCA AGAC TCCCCA CGGTGT TGG TCCTA GACC TTTTGT CGATGAT
GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG
CAGG CA AG ACT GGAGG AGGAGGA ACTGG GGTGT AAGG CC AGGG TGT GGC CCTGCG GCG GAC GT AAGA AGGGGA GATGG TCCAGA CCTT TGGAG
GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG
GAG ATTG CGC TGTCTC AA AAGGATA GG CTG CGGCTGC GCTGCA GGGC CGC CGCT GACAGGCA GTTGGAT AAGAG AGGGGCT GCAGGT GCAGTGCT GAGGACTT AGGGCC
GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG
13 13 13 13 13 14 14 14 14 14 14 14 14 15 15 15 16 16 18 18 18
Position relative to Transcription start site (TSS). CD signature obtained for quadruplex (10 µM) in 10 mM sodium cacodylate buffer, pH 7.4 with 100 mM KCl. The respective signature of quadruplex is defined as 1) Parallel suggests predominant parallel population. 1 2) Parallel suggests predominant parallel and some antiparallel population. 3) Antiparallel suggests predominant antiparallel population. 1 4) Antiparallel suggests predominant antiparallel and a small parallel population. 5) Mixed suggests existence of both parallel and antiparallel population. b
Mixed Mixed Mixed Parallel 1 Parallel 1 Parallel 1 Parallel Antiparallel1 Mixed Mixed Parallel 1 Antiparallel1 Mixed Mixed Mixed Parallel 1 Parallel 1 Mixed Mixed Mixed Mixed
SI Table 2: The quadruplex forming sequences which were excluded from the analysis.
Oligo Name
Gene
Q1 Q2 Q3 Q4 Q5 Q6 Q7 Q8 Q9 Q10 Q11 Q12 Q13 Q14 Q15 Q16 Q17 Q18
USP41 KLHL14 GGTLA1 TACC3 GLI1 TCEB3C JUNB TAL1 ETV6 AKT1 FLI1 FLI1 MYEOV THPO YES1 VEGF FES CSF1R
Position relative to TSSa -43 -42 -1259 -156 -382 -1190 -1695 -314 -63 -39 -213 -205 -751 -1688 -120 -85 -247 -1105
Total
Sequence (5′-3′)
GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG
A G T G GC GGC T G A GC GGTGT G CGGTGA GGAGG TTCGC GCGGGCC CGCTGCC GTATCCT
GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG
A G T A G G GC GCGGT CGGG CGC G GA TAAA AGGGGA TGGGAA GGCG CCCTG CAAAA
GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG
A G T A G G GCC G CGG GCG GA AAGACGA AGGAGGA ACCAGG ACGCCGC TCCCGGC CCTGCG ACAAAGA
These sequences did not give characteristic quadruplex UV melting/cooling profile at 295 nm in 10 mM sodium cacodylate pH 7.4, 100 mM KCl, and were excluded from further experiments.
GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG GGG
3 3 3 3 4 5 6 7 8 8 8 10 17 17 18 18 18 19
SI Table 3 A: Thermodynamic profile obtained for sequences with loop length of 5-9.
Oligo Name
Gene
Total
ΔΗ kcal/mol
Q1 Q2 Q3 Q4 Q5 Q6 Q7 Q8 Q9 Q10 Q11 Q12 Q13 Q14 Q15 Q16 Q17 Q18 Q19 Q20
VEGF WNT 3 WNT5A PDGFB JUNB TAL1 THPO FOS PIM1 FGF 3 FES VAV1 IGF2 FOSL1 TAL1 AKT2 THRA FBXW 7 HRAS WNT1
5 5 5 6 6 6 6 7 7 7 7 7 8 8 8 8 8 8 9 9
-51.5 ±0.8 -101.5 ± 5.0 -39.6 ± 2.5 -67.0 ± 6.0 nd nd -37.0 ± 4.0 -30.5 ± 1.4 -69.3 ± 6.0 -22.3 ± 0.5 -58.0 ± 0.4 -19.0 ± 1.2 -28.0 ± 2.4 -37.0 ± 0.6 -44.0 ± 1.0 nd nd -30.4 ± 0.8 -28.8 ± 2.0 -24.0 ± 3.0
Quadruplex ΔS cal/mol/K -142.1± 2.2 -281.7 ± 10.0 -119.0 ± 5.8 -188.0 ± 9.0
ΔG37 kcal/mol -4.6 ± 0.1 -17.6 ± 2.0 -2.7± 0.7 -12.4 ± 3.2
-100.0± 10.0 -87.7 ± 4.1 -194 ± 25.5 -66.0 ± 1.2 -175.6 ± 1.2 -54.0 ± 3.3 -82.0 ± 7.7 -104 ± 1.7 -125.0± 2.7
-3.3 ± 1.0 -2.5 ± 0.1 -9.8 ±1.2 -1.3 ± 0.1 -2.6 ± 0.1 -1.6 ± 0.2 -1.0 ± 0.1 -3.5 ± 0.1 -6.2 ± 0.2
-87.0 ± 2.2 -91.2 ± 5.0 -74.6 ± 7.5
-3.6 ± 0.2 -1.5 ± 0.5 -0.9± 0.7
The data was simulated with Gaussian distribution of random error and the data was then fit to nonlinear regression fitting followed by Monte Carlo analysis performed by fitting 1000 simulated data sets. (Details provided in materials and method)
SI Table 3 B: Thermodynamic profile obtained for sequences with loop length of 10-14. Oligo Name
Gene
Total
Q21 Q22 Q23 Q24 Q25 Q26 Q27 Q28 Q29 Q30 Q31 Q32 Q33 Q34 Q35 Q36 Q37 Q38 Q39 Q40 Q41 Q42 Q43 Q44 Q45 Q46 Q47 Q48 Q49 Q50 Q51 Q52 Q53 Q54
c-MYC NFKB 2 GLI 1 HCK HCK YES 1 AKT 1 BCL 2 NOTCH 1 c-KIT JUNB SRC VAV 2 AKT 2 MYB ROS 1 TAL 1 WNT1 CTTN AXL FGF 3 MAF MPL MYB ABL 1 MYEOV VEGF FLI 1 FGR WNT 3 CSF 1R THRA FES REL
10 10 10 10 10 11 11 11 11 11 11 11 11 11 11 11 12 12 12 12 12 13 13 13 13 13 13 14 14 14 14 14 14 14
ΔΗ kcal/mol -33.5 ± 0.3 -43.0 ± 1.4 -44.5 ± 2.7 -38.8 ± 1.7 nd nd nd nd nd -30.0± 0.3 nd nd nd -72.3 ± 3.3 -64.5 ± 3.8 -69.0 ± 3.0 nd -36.6 ± 1.0 nd -44.4 ± 0.7 -38.0 ± 0.8 nd nd nd -74.0 ± 5.0 -110.0 ± 8.5 -34.0 ± 1.3 -24.0 ± 1.8 -33.0 ± 0.8 nd nd -51.0 ± 2.1 -34.0 ± 0.3 nd
Quadruplex ΔS cal/mol/K -97.4 ± 1.0 -122.0 ± 4.1 -127.0 ± 7.8 -120.0 ± 3.6
ΔG37 kcal/mol -2.5 ± 0.1 -3.72 ± 0.1 -4.6 ± 0.3 -1.5 ± 0.6
-82.0± 1.0
-4.00± 0.1
-197.0± 8.4 -175.0± 10.0 -194.0± 8.0
-5.0 ± 0.7 -4.6 ± 0.7 -5.2 ± 0.5
-1043.0± 3.0
-3.3 ± 0.1
-129.0 ± 2.2 -112.0 ± 2.5
-5.3 ± 0.1 -1.7 ± 0.1
-210.5 ± 14.0 -330.0 ± 25.0 -102.6 ± 3.8 -70.0 ± 5.0 -100.0 ± 2.3
-5.2 ± 0.7 -4.2 ± 0.8 -2.0 ± 0.1 -4.8 ± 0.3 -2.1 ± 0.1
-142.0 ± 6.2 -99.0 ± 0.8
-5.6 ± 0.2 -3.5 ± 0.1
The data was simulated with Gaussian distribution of random error and the data was then fit to nonlinear regression fitting followed by Monte Carlo analysis performed by fitting 1000 simulated data sets. (Details provided in materials and method)
SI Table 3 C: Thermodynamic profile obtained for sequences with loop length of 15-18. Oligo Name
Gene
Total
Q55 Q56 Q57 Q58 Q59 Q60 Q61 Q62
FGF 4 BCL 3 KRAS THPO RALB EGFR VAV 1 FGF 6
15 15 15 16 16 18 18 18
ΔΗ kcal/mol nd nd -45.0 ± 2.0 -70.5 ± 3.8 nd nd nd nd
Quadruplex ΔS cal/mol/K
ΔG37 kcal/mol
-130.0 ± 6.0 -195.0 ± 10.0
-1.2 ± 0.2 -4.3 ± 0.7
The data was simulated with Gaussian distribution of random error and the data was then fit to nonlinear regression fitting followed by Monte Carlo analysis performed by fitting 1000 simulated data sets. (Details provided in materials and method)
A 1 2 3 4 5 6 7 8 9 10 11 12 13
Lane 1: T25; Lane 2: T33; Lane 3: T38; Lane 4: Q1; Lane 5: Q2; Lane 6: Q3; Lane 7: Q4; Lane 8: Q5; Lane 9: Q6; Lane 10: Q7; Lane 11: Q8; Lane 12: Q9; Lane 13: Q10.
B
1
2
3 4 5 6 7 8 9 10 11 12 13
Lane 1: T25; Lane 2: T33; Lane 3: T38; Lane 4: Q11; Lane 5: Q12; Lane 6: Q13; Lane 7: Q14; Lane 8: Q15; Lane 9: Q16; Lane 10: Q17; Lane 11: Q18; Lane 12: Q19; Lane 13: Q20.
C 1
2
3 4
5 6
7
8
9 10 11 12 13
Lane 1: T25; Lane 2: T33; Lane 3: T38; Lane 4: Q21; Lane 5: Q22; Lane 6: Q23; Lane 7: Q24; Lane 8: Q30; Lane 9: Q34;Lane 10: Q35 (L11); Lane 11: Q36; Lane 12: Q38; Lane 13: Q40. D
1
2
3
4 5
6
7 8
9 10 11 12 13
Lane 1: T25; Lane 2: T33; Lane 3: T38; Lane 4: Q41; Lane 5: Q45; Lane 6: Q46; Lane 7: Q47; Lane 8: Q48; Lane 9: Q49; Lane 10: Q52; Lane 11: Q53; Lane 12: Q57; Lane 13: Q58. SI Figure 1 (A-D): Gel electrophoresis for 40 sequences with 30 μM concentration in 100 mM KCl Buffer, pH 7.4 at 4°C with marker as dT25, dT33 and dT38.
CD spectra and UV cooling curves of the sequences used in this study in 10 mM sodium cacodylate, pH 7.4, 100 mM KCl. a Raw data for UV melting study along with their non-linear least square curve fittings is represented for selected 36 sequences. (Q1) VEGF
0.21
12
0.20 Abs 295 nm
θ, mdeg
9 6 3 0
0.18 0.17
-3 -6
0.19
0.16 15 30 45 60 75 90 0 Temp, C
225 250 275 300 325 wavelength, nm (Q2) WNT 3
0.21
12
0.20 Abs 295 nm
θ, mdeg
9 6 3 0
0.18 0.17
-3 -6
0.19
0.16 15 30 45 60 75 90 105 0 Temp, C
225 250 275 300 325 wavelength, nm
0.18
12 9 6 3 0 -3 -6
Abs 295 nm
θ, mdeg
(Q3) WNT 5A
0.16 0.14 0.12 0.10
225 250 275 300 325 wavelength, nm
0.08 15 30 45 60 75 90 105 0 Temp, C
(Q 4) PDGFB
0.21 9 6
Abs 295 nm
θ, mdeg
12
3 0 -3 -6
0.20 0.19 0.18 0.17 15 30 45 60 75 90 105 0 Temp, C
225 250 275 300 325 wavelength, nm
12 9 6 3 0 -3 -6
0.18 Abs 295 nm
θ, mdeg
(Q5) JUNB
0.17 0.16 0.15 15 30 45 60 75 90 105 0 Temp, C
225 250 275 300 325 wavelength, nm (Q6) TALI
0.18 Abs 295 nm
θ, mdeg
12 9 6 3 0 -3 -6
0.17 0.16 0.15 15 30 45 60 75 90 105 0 Temp, C
225 250 275 300 325 wavelength, nm
12
0.192
9 6
0.186
Abs 295 nm
θ, mdeg
(Q7) THPO
3 0 -3 -6
0.180 0.174 0.168
225 250 275 300 325 wavelength, nm
0.162 15 30 45 60 75 90 105 0 Temp, C
(Q8) FOS
0 -3 -6
Abs 295 nm
θ, mdeg
0.22 12 9 6 3
0.21 0.20 0.19 0.18
225 250 275 300 325 wavelength, nm
15 30 45 60 75 90 105 0 Temp, C
12 9 6 3 0 -3 -6
0.20 Abs 295 nm
θ, mdeg
(Q9) PIM 1
0.19 0.18
225 250 275 300 325 wavelength, nm
15 30 45 60 75 90 105 0 Temp, C
(Q10) FGF 3
0 -3 -6
0.20 Abs 295 nm
θ, mdeg
0.22 12 9 6 3
0.18 0.16 0.14 0.12 15 30 45 60 75 90 105 0 Temp, C
225 250 275 300 325 wavelength, nm
12 9 6 3 0 -3 -6
Abs 295 nm
θ, mdeg
(Q11) FES
225 250 275 300 325 wavelength, nm
0.21 0.18 0.15 0.12 0.09 0.06 0.03 0.00 15 30 45 60 75 90 105 0 Temp, C
0.225 Abs 295 nm
θ, mdeg
(Q12) VAV 1
15 12 9 6 3 0 -3 -6
0.210 0.195 0.180 15 30 45 60 75 90 105 0 Temp, C
225 250 275 300 325 wavelength, nm (Q13) IGF 2
12
0.185 Abs 295 nm
θ, mdeg
9 6 3 0
0.175 0.170
-3 -6
0.180
225 250 275 300 325 wavelength, nm
0.165
15 30 45 60 75 90 0 Temp, C
0.20
12 9 6
Abs 295 nm
θ, mdeg
(Q14) FOSL 1
3 0 -3 -6
0.19 0.18 0.17 0.16 0.15 0.14 15 30 45 60 75 90 105 0 Temp, C
225 250 275 300 325 wavelength, nm
16 12 8 4 0 -4 -8
Abs 295 nm
θ, mdeg
(Q15) TAL 1
225 250 275 300 325 wavelength, nm
0.23 0.22 0.21 0.20 0.19 0.18 0.17 0.16 0.15 45 60 75 90 105 0 Temp, C
(Q16) AKT 2
12
0.20 Abs 295 nm
θ, mdeg
8 4 0 -4
0.19 0.18 0.17
225 250 275 300 325 wavelength, nm
15 30 45 60 75 90 105 0 Temp, C
12
0.20
8
0.19
Abs 295 nm
θ, mdeg
(Q17) THRA
4 0
0.18 0.17 0.16 0.15
-4
0.14
225 250 275 300 325 wavelength, nm
45
60 75 90 105 0 Temp, C
0.19 0.18 Abs 295 nm
θ, mdeg
(Q18) FBXW 7
18 15 12 9 6 3 0 -3 -6 -9
0.17 0.16 0.15 0.14 0.13 15 30 45 60 75 90 105 0 Temp, C
225 250 275 300 325 wavelength, nm (Q19) HRAS
0.175
12 Abs 295 nm
θ, mdeg
8 4 0
0.170 0.165 0.160
-4 225 250 275 300 325 wavelength, nm
0.155 15 30 45 60 75 90 105 0 Temp, C
(Q 20) WNT 1
0.21 Abs 295 nm
θ, mdeg
12 8 4 0
0.20 0.19 0.18 0.17
-4
0.16 225 250 275 300 325 wavelength, nm
15 30 45 60 75 90 0 Temp, C
(Q21) c-MYC
Abs 295 nm
θ, mdeg
12 8 4 0 -4 225 250 275 300 325 wavelength, nm
0.20 0.18 0.16 0.14 0.12 0.10 0.08 0.06
30 45 60 75 90 105 0 Temp, C
(Q22) NFKB 2
0.180 Abs 295 nm
θ, mdeg
12 8 4 0
0.165 0.150 0.135
-4 225 250 275 300 325 wavelength, nm
30
45 60 75 90 105 0 Temp, C
(Q23) GLI 1
12 0.240 Abs 295 nm
θ, mdeg
8 4 0
0.225
-4 -8 225 250 275 300 325 wavelength, nm
0.210 15 30 45 60 75 90 105 0 Temp, C
(Q24) HCK
12
0.180 Abs 295 nm
θ, mdeg
8 4 0 -4
0.165 0.150 0.135
-8
0.120 15 30 45 60 75 90 105 0 Temp, C
225 250 275 300 325 wavelength, nm
0.22
16 12 8 4 0 -4 -8
0.20 Abs 295 nm
θ, mdeg
(Q25) HCK
0.18 0.16 0.14 0.12 0.10 15 30 45 60 75 90 105 0 Temp, C
225 250 275 300 325 wavelength, nm
0.22
16 12 8 4 0 -4 -8
0.20 Abs 295 nm
θ, mdeg
(Q26) YES 1
0.18 0.16 0.14 0.12 0.10 15 30 45 60 75 90 105 0 Temp, C
225 250 275 300 325 wavelength, nm (Q27) AKT 1
12
0.210 Abs 295 nm
θ, mdeg
8 4 0
0.200 0.195
-4 -8
0.205
225 250 275 300 325 wavelength, nm
0.190 15 30 45 60 75 90 0 Temp, C
0.213
8
0.210 Abs 295 nm
θ, mdeg
(Q28) BCL 2
12
4 0 -4
0.207 0.204 0.201
-8
0.198 225 250 275 300 325 wavelength, nm
30
45 60 75 0 Temp, C
90
(Q29) NOTCH 1
12 0.180 Abs 295 nm
θ, mdeg
8 4 0
0.177 0.174
-4
0.171
-8
0.168 225 250 275 300 325 wavelength, nm
15 30 45 60 75 90 105 0 Temp, C
(Q30) c-KIT
0.21
8
Abs 295 nm
θ, mdeg
12
4 0
0.15 0.12 0.09
-4 -8
0.18
0.06 15 30 45 60 75 90 105 0 Temp, C
225 250 275 300 325 wavelength, nm (Q31) JUNB
0.21
8
Abs 295 nm
θ, mdeg
12
4 0
0.18 0.15
-4 -8
225 250 275 300 325 wavelength, nm
0.12 15 30 45 60 75 90 105 0 Temp, C
(Q32) SRC
0.20
8
Abs 295 nm
θ, mdeg
12
4 0 -4 -8
0.18 0.16 0.14 0.12
225 250 275 300 325 wavelength, nm
15 30 45 60 75 90 0 Temp, C
(Q33) VAV 2
0.19
12
0.18 Abs 295 nm
θ, mdeg
8 4 0 -4
0.16 0.15
-8 -12
0.17
0.14 15 30 45 60 75 90 105 0 Temp, C
225 250 275 300 325 wavelength, nm (Q34) AKT 2
0.21 0.20
9 6
Abs 295 nm
θ, mdeg
12
3 0 -3 -6
0.19 0.18 0.17 0.16 0.15
225 250 275 300 325 wavelength, nm
15 30 45 60 75 90 105 0 Temp, C
(Q35) MYB
12
0.225 Abs 295 nm
θ, mdeg
9 6 3 0
0.215 0.210
-3 -6
0.220
225 250 275 300 325 wavelength, nm
0.205 30
45 60 75 90 105 0 Temp, C
16 12 8 4 0 -4 -8 -12
Abs 295 nm
θ, mdeg
(Q36) ROS 1
225 250 275 300 325 wavelength, nm
0.21 0.20 0.19 0.18 0.17 0.16 0.15 0.14 15 30 45 60 75 90 105 0 Temp, C
0.16
9
0.15 Abs 295 nm
θ, mdeg
(Q37) TAL 1
12 6 3 0
0.13 0.12
-3 -6
0.14
0.11 15 30 45 60 75 90 105 0 Temp, C
225 250 275 300 325 wavelength, nm
0.18 Abs 295 nm
θ, mdeg
(Q38) WNT 1
15 12 9 6 3 0 -3 -6
0.17 0.16 0.15 15 30 45 60 75 90 105 0 Temp, C
225 250 275 300 325 wavelength, nm
0.19 Abs 295 nm
θ, mdeg
(Q39) CTTN
15 12 9 6 3 0 -3 -6
0.18 0.17 0.16 0.15
225 250 275 300 325 wavelength, nm
15 30 45 60 75 90 105 0 Temp, C
0.20 Abs 295 nm
θ, mdeg
(Q40) AXL
18 15 12 9 6 3 0 -3 -6 -9
0.18 0.16 0.14 0.12 15 30 45 60 75 90 105 0 Temp, C
225 250 275 300 325 wavelength, nm
0.27 Abs 295 nm
θ, mdeg
(Q41) FGF 3
18 15 12 9 6 3 0 -3 -6 -9
0.24 0.21 0.18 0.15 0.12 15 30 45 60 75 90 105 0 Temp, C
225 250 275 300 325 wavelength, nm
0.21 0.18 Abs 295 nm
θ, mdeg
(Q42) MAF
18 15 12 9 6 3 0 -3 -6 -9
0.15 0.12 0.09 0.06 15 30 45 60 75 90 105 0 Temp, C
225 250 275 300 325 wavelength, nm
0.21 Abs 295 nm
θ, mdeg
(Q43) MPL
18 15 12 9 6 3 0 -3 -6 -9
0.18 0.15 0.12 0.09
225 250 275 300 325 wavelength, nm
0.06 15 30 45 60 75 90 105 0 Temp, C
(Q44) MYB
12
0.210 Abs 295 nm
θ, mdeg
9 6 3 0
0.195 0.180
-3 -6
0.165 15 30 45 60 75 90 105 0 Temp, C
225 250 275 300 325 wavelength, nm (Q45) ABL 1
15
0.200 Abs 295 nm
θ, mdeg
10 5 0 -5
0.195 0.190 0.185
-10
0.180
-15
0.175 225 250 275 300 325 wavelength, nm
30 45 60 75 90 105 0 Temp, C
(Q46) MYEOV
0.22 0.21 Abs 295 nm
θ, mdeg
15 10 5 0
0.20 0.19 0.18 0.17
-5 225 250 275 300 325 wavelength, nm
30
45 60 75 0 Temp, C
90
(Q47) VEGF
0.18 Abs 295 nm
θ, mdeg
15 10 5
0.17 0.16 0.15
0
0.14
-5
0.13 225 250 275 300 325 wavelength, nm
15 30 45 60 75 90 105 0 Temp, C
(Q48) FLI 1
0.240 Abs 295 nm
θ, mdeg
15 10 5 0
0.232 0.224
-5 225 250 275 300 325 wavelength, nm
15 30 45 60 75 90 105 0 Temp, C
(Q49) FGR
Abs 295 nm
θ, mdeg
15 10 5 0 -5 225 250 275 300 325 wavelength, nm
0.21 0.20 0.19 0.18 0.17 0.16 0.15 15 30 45 60 75 90 105 0 Temp, C
0.27
15
0.26 Abs 295 nm
θ, mdeg
(Q50) WNT 3
20 10 5 0 -5 -10
0.25 0.24 0.23 0.22 0.21 15 30 45 60 75 90 105 0 Temp, C
225 250 275 300 325 wavelength, nm
0.21
15
0.20 Abs 295 nm
θ, mdeg
(Q51) CSF 1R
20 10 5 0
0.18 0.17 0.16
-5 -10
0.19
0.15 225 250 275 300 325 wavelength, nm
15 30 45 60 75 90 105 0 Temp, C
0.25 0.24 Abs 295 nm
θ, mdeg
(Q52) THRA
20 15 10 5 0 -5 -10 -15
0.23 0.22 0.21 0.20
225 250 275 300 325 wavelength, nm
30 45 60 75 90 105 0 Temp, C
(Q53) FES
0.20 Abs 295 nm
θ, mdeg
10 5 0
0.18 0.16 0.14 0.12
-5
0.10 15 30 45 60 75 90 105 0 Temp, C
225 250 275 300 325 wavelength, nm (Q54) REL
15
0.20 Abs 295 nm
θ, mdeg
10 5 0
0.18 0.16 0.14
-5 225 250 275 300 325 wavelength, nm
15 30 45 60 75 90 105 0 Temp, C
(Q55) FGF 4
0.21 Abs 295 nm
θ, mdeg
10 5 0
0.20 0.19 0.18
-5 225 250 275 300 325 wavelength, nm
0.17
30
45 60 75 0 Temp, C
90
(Q56) BCL 3
0.175
15 Abs 295 nm
θ, mdeg
10 5 0
0.170 0.165
-5 0.160
225 250 275 300 325 wavelength, nm
30
45 60 75 0 Temp, C
90
(Q57) KRAS
0.20
10
Abs 295 nm
θ, mdeg
15
5 0 -5 -10
0.18 0.16
225 250 275 300 325 wavelength, nm
15 30 45 60 75 90 105 0 Temp, C
(Q58) THPO
0.255
10
Abs 295 nm
θ, mdeg
15
5 0 -5 -10
0.240
0.225 225 250 275 300 325 wavelength, nm
15 30 45 60 75 90 105 0 Temp, C
(Q59) RALB
0.210 10
Abs 295 nm
θ, mdeg
15
5 0
0.195 0.180
-5 -10
225 250 275 300 325 wavelength, nm
0.165 15 30 45 60 75 90 0 Temp, C
(Q60) EGFR
0.21 15
0.20 Abs 295 nm
θ, mdeg
10 5 0 -5
0.19 0.18 0.17 0.16
-10
0.15 15 30 45 60 75 90 105 0 Temp, C
225 250 275 300 325 wavelength, nm (Q61) VAV 1
0.195
10
Abs 295 nm
θ, mdeg
15
5 0 -5 -10
0.180 0.165 0.150
225 250 275 300 325 wavelength, nm
15 30 45 60 75 90 0 Temp, C
(Q62) FGF 6
0.18 10
Abs 295 nm
θ, mdeg
15 5 0 -5
0.17 0.16 0.15
-10 225 250 275 300 325 wavelength, nm
15 30 45 60 75 90 105 0 Temp, C
