Table S1 List of primer combinations for TRα1 ... - Semantic Scholar

Table S1 List of primer combinations for TRα1 constructs with respective inserts cloned Primers

a,b

Pr-1, 2 Pr-3, 4 Pr-5, 6 Pr-7, 8 Pr-1, 4 Pr-33, 34 Pr-37, 35 Pr-36, 34 Pr-33, 35 Pr-36, 46 Pr-47, 48 Pr-37, 49 Pr-37, 50 Pr-37, 51 Pr-37, 52 Pr-57, 58 Pr-67, 68 Pr-77, 78 Pr-79, 80 Pr-85, 86 Pr-87, 88 Pr 91, 92 Pr-93, 94 Pr-147,148 Pr-149,150 Pr-153,154 Pr-155,156 Pr-95, 96 Pr-99, 100 Pr-101, 102 Pr-103, 104 Pr-107, 108 Pr-109, 110 Pr-111, 112 Pr-114, 115 Pr-116, 117 Pr-133, 134 Pr-135, 136 Pr-137, 138 Pr-135, 136 Pr-137, 138 Pr-139, 140 Pr-141, 142

Insert/s A/B domain DBD (1 amino acid less) Hinge domain LBD A/B-DBD DBD-Hinge Hinge-LBD A/B-DBD-Hinge DBD-Hinge-LBD A/B-LBD A/B-Hinge-LBD Hinge-LBD∆408-410 Hinge-LBD∆393-410 Hinge-LBD∆268-410 Hinge-LBD∆206-410 NESH12 - Hinge NLS1 NESH12-F401A-Hinge NESH12-F397A,F401A-Hinge NESH12-P398G-Hinge NESH3H6-Hinge NESH3-Hinge NESH6-Hinge NESH12-NESH3-Hinge NESH12-NESH6-Hinge NESH12-NESH12F401A-Hinge NESH12-NESH12P453T-Hinge NESH12L400S-Hinge NESH12L4A-Hinge NESH12L400P-Hinge NESH12P399T-Hinge NESH12F401G-Hinge NESH12F401P-Hinge NESH12F401N-Hinge NESH12F401P,F405P-Hinge NESH12F401P,F405N-Hinge 2(NESH12 )–Hinge NESH12-WT+F401P-Hinge NESH12-WT+F401P,F405N-Hinge NESH12-F401P+F401P-Hinge “ NESH12-F401P+ F401P,F405N -Hinge hTRb1-A/BD hTRb1-DBD

c

d

RE Sites

Source Vector

Kpn I, Xba I “ “ “ “ “ “ “ “ Hind III, Kpn I “ “ “ “ “ “ “ “ Xho I, Hind III Hind III, Kpn I “ “I “ “ “ “ Xho I, Hind III “ “ “ “ “ “ “ “ “ “ “ “ “ Hind II Kpn I “ “ “

EG3 “ “ “ “ “ “ “ “ EG3-LBD EG3-Hinge-LBD EG3 “ “ “ EG3-Hinge EG3 EG3-Hinge “ “ “ “ “ “ EG3 NESH12-Hinge “ “ “ “ “ “ “ “ “ “ “ “ “ EG3-NESH12-Hinge “ “ EG3-NESH12-F401P-Hinge

“ “ “ “ “ “ “ “ “ “ “ “ “ “ “ “ “ “ Hind III, Kpn I “ “ “ “ “ “ “ Kpn I, Xba I “ “

“ EG3 “

NLS-2 Oligopeptide Mutations Pr-31, 32 Pr-38, 39 Pr-40, 41 Pr-42, 43 Pr-44, 45 Pr-167, 168 Pr-169, 170 Pr-171, 172

NLS2-WT NLS2-K29A NLS2-P22L NLS2-P22L-R26H NLS2-5K/R NLS2*-P22A NLS2*-P22AG24A NLS2* P22AD23A

Hind III, “ “ “ “ “ “

Kpn I “ “ “ “ “ “ “

EG3 “ “ “ “ “ “ “

“

A/B Domain Mutagenesis Primers and Respective Mutations Pr-11, 12

R26H

None

EG3-A/B Domain

Pr-13, 14 Pr-15 , 16 Pr-17, 18 Pr-23, 24 Pr-25, 26 Pr-163, 164 Pr-165, 166 Pr-159, 160 Pr-161, 162

K27A,R28A P22A K25A K25A, R26A R28A K29A P22—G24A vErbA A/B domain cTRa1 A/B domain

“ “ “ “ “ “ “ Kpn I, Xba I “

“ “ “ “ “ “ “ EG3 “

None None “

GFP-TRα1 “ GFP-TRβ1

TRα1 and TR β1 whole protein mutations Pr-11, 12 Pr-174, 175 Pr-182, 183 a

TRα1 R26H TRα1 F401A TRβ1 F401A

Sequence of individual primers or oligonucleotides mentioned in this table are presented in Table S2. The first primer in a pair is the forward primer and the second is the reverse primer. c RE sites, restriction endonuclease sites. d EG3, EGFPC1 based GFP-GST-GFP vector. b

Table S2 List of sequences of primers used in the current study Primer Name

Sequence

Pr-1

ggggtaccatggaacagaagccaagca

Pr-2

gctctagactgctcgtctttgtccaggt

Pr-3

ggggtacctgtgtcgtgtgtggggac

Pr-4

gctctagatagaaccaggtccatggcc

Pr-5

ggggtaccgattcaaagcgggtggc

Pr-6

gctctagatcgcctctgtttccaatg

Pr-7

ggggtaccaaattcctgccggatgac

Pr-8

gctctagagacttcctgatcctcaaagacct

Pr-11

aggtcaccagatggaaagcataaaaggaagaacggccaat

Pr-12

attggccgttcttccttttatgctttccatctggtgacct

Pr-13

ggtcaccagatggaaagcgagcagcgaagaacggccaatgtcccc

Pr-14

ggggacattggccgttcttcgctgctcgctttccatctggtgacc

Pr-15

gaacagtgccaggtcagcagatggaaagcgaaa

Pr-16

tttcgctttccatctgctgacctggcactgttc

Pr-17

cagtgccaggtcaccagatggagcgcgaaaaaggaagaa

Pr-18

ttcttcctttttcgcgctccatctggtgacctggcactg

Pr-23

gtgccaggtcaccagatggagcggcaaaaaggaagaacggccaat

Pr-24

attggccgttcttcctttttgccgctccatctggtgacctggcac

Pr-25

accagatggaaagcgaaaagcgaagaacggccaatgtcc

Pr-26

ggacattggccgttcttcgcttttcgctttccatctggt

Pr-31

cccaagcttcgccagatggaaagcgaaaaaggaagggtacccc

Pr-32

ggggtacccttcctttttcgctttccatctggcgaagcttggg

Pr-33

cccaagcttcgtgtgtcgtgtgtggggac

Pr-34

ggggtacctcgcctctgtttccaatgg

Pr-35

ggggtaccgacttcctgatcctcaaagacct

Pr-36

cccaagcttcgatggaacagaagccaagcaag

Pr-37

cccaagcttcggattcaaagcgggtggcc

Pr-38

cccaagcttcgccagatggaaagcgaaaaagggcgggtacccc

Pr-39

ggggtacccgccctttttcgctttccatctggcgaagcttggg

Pr-40

cccaagcttcgctagatggaaagcgaaaaaggaagggtacccc

Pr-41

ggggtacccttcctttttcgctttccatctagcgaagcttggg

Pr-42

cccaagcttcgctagatggaaagcacaaaaggaagggtaccc

Pr-43

ggggtacccttccttttgtgctttccatctagcgaagcttggg

Pr-44

cccaagcttcggcagcgaagcgaaaaaggaaggcgggtacccc

Pr-45

ggggtacccgccttcctttttcgcttcgctgccgaagcttggg

Pr-46

ggggtaccctgctcgtctttgtccaggt

Pr-47

ccgctcgagctatggaacagaagccaagcaag

Pr-48

cccaagcttgctgctcgtctttgtccaggt

Pr-49

ggggtaccatcctcaaagacctccaggaa

Pr-50

ggggtaccgcactcgactttcatgtgga

Pr-51

ggggtaccgtcatagcggacagctgcc

Pr-52

ggggtacccggcatggagacaataggtg

Pr-57

ccgctcgagctgtcgagtgccccaccgaactcttccccccactcttcctggaggtctttgaggatcaagcttccc

Pr-58

gggaagcttgatcctcaaagacctccaggaagagtggggggaagagttcggtggggcactcgacagctcgagcgg

Pr-67

ccgctcgagctaagcgggtggccaaacgcaagctgattgagcagaaccgggagaggaggcgaaagcaagcttccc

Pr-68

gggaagcttgctttcgcctcctctcccggttctgctcaatcagcttgcgtttggccacccgcttagctcgagcgg

Pr-77

ccgctcgagctgtcgagtgccccaccgaactcttccccccactcgcactggaggtctttgaggatcaagcttccc

Pr-78

gggaagcttgatcctcaaagacctccagtgcgagtggggggaagagttcggtggggcactcgacagctcgagcgg

Pr-79

ccgctcgagctgtcgagtgccccaccgaactcgcacccccactcgcactggaggtctttgaggatcaagcttccc

Pr-80

gggaagcttgatcctcaaagacctccagtgcgagtgggggtgcgagttcggtggggcactcgacagctcgagcgg

Pr-85

ccgctcgagctgtcgagtgccccaccgaactcttcggaccactcttcctggaggtctttgaggatcaagcttccc

Pr-86

gggaagcttgatcctcaaagacctccaggaagagtggtccgaagagttcggtggggcactcgacagctcgagcgg

Pr-87

ccgctcgagctaaggtagacctagaggccttcag

Pr-88

gggaagcttggacagctgcccgcagggac

Pr-91

ccgctcgagctaaggtagacctagaggccttca

Pr-92

gggaagcttgaggcagctcggagaacatgg

Pr-93

ccgctcgagctcccatgttctccgagctgcc

Pr-94

gggaagcttggacagctgcccgcagggac

Pr-95

ccgctcgagctgtcgagtgccccaccgaactcttccccccatccttcctggaggtctttgaggatcaagcttccc

Pr-96

gggaagcttgatcctcaaagacctccaggaaggatggggggaagagttcggtggggcactcgacagctcgagcgg

Pr-99

ccgctcgagctgtcgagtgccccaccgaactcttccccccagcagctctggaggctgcagaggatcaagcttccc

Pr-100

gggaagcttgatcctctgcagcctccagagctgctggggggaagagttcggtggggcactcgacagctcgagcgg

Pr-101

ccgctcgagctgtcgagtgccccaccgaactcttccccccaccgttcctggaggtctttgaggatcaagcttccc

Pr-102

gggaagcttgatcctcaaagacctccaggaacggtggggggaagagttcggtggggcactcgacagctcgagcgg

Pr-103

ccgctcgagctgtcgagtgccccaccgaactcttccccacactcttcctggaggtctttgaggatcaagcttccc

Pr-104

gggaagcttgatcctcaaagacctccaggaagagtgtggggaagagttcggtggggcactcgacagctcgagcgg

Pr-107

ccgctcgagctgtcgagtgccccaccgaactcttccccccactcGGActggaggtctttgaggatcaagcttccc

Pr-108

gggaagcttgatcctcaaagacctccagtccgagtggggggaagagttcggtggggcactcgacagctcgagcgg

Pr-109

ccgctcgagctgtcgagtgccccaccgaactcttccccccactcCCTctggaggtctttgaggatcaagcttccc

Pr-110

gggaagcttgatcctcaaagacctccagagggagtggggggaagagttcggtggggcactcgacagctcgagcgg

Pr-111

ccgctcgagctgtcgagtgccccaccgaactcttccccccactcAAcctggaggtctttgaggatcaagcttccc

Pr-112

gggaagcttgatcctcaaagacctccaggttgagtggggggaagagttcggtggggcactcgacagctcgagcgg

Pr-114

ccgctcgagctgtcgagtgccccaccgaactcttccccccactccctctggaggtccctgaggatcaagcttccc

Pr-115

gggaagcttgatcctcagggacctccagagggagtggggggaagagttcggtggggcactcgacagctcgagcgg

Pr-116

ccgctcgagctgtcgagtgccccaccgaactcttccccccactcCCTctggaggtcaacgaggatcaagcttccc

Pr-117

gggaagcttgatcctcgttgacctccagagggagtggggggaagagttcggtggggcactcgacagctcgagcgg

Pr-133

cccaagcttcggtcgagtgccccaccgaactcttcccTccactcttcctggaggtctttgaggatggtacccc

Pr-134

ggggtaccatcctcaaagacctccaggaagagtggAgggaagagttcggtggggcactcgaccgaagcttggg

Pr-135

cccaagcttcggtcgagtgccccaccgaactcttccccccactccctctggaggtctttgaggatggtacccc

Pr-136

ggggtaccatcctcaaagacctccagagggagtggggggaagagttcggtggggcactcgaccgaagcttggg

Pr-137

cccaagcttcggtcgagtgccccaccgaactcttccccccactccctctggaggtcaacgaggatggtacccc

Pr-138

ggggtaccatcctcgttgacctccagagggagtggggggaagagttcggtggggcactcgaccgaagcttggg

Pr-139

ggggtaccatgactcccaacagtatgacag

Pr-140

gctctagagagctcgtccttgtctaagta

Pr-141

ggggtacctgtgtagtgtgtggtgacaaag

Pr-142

gctctagaatccagcaccaaatctgttgc

Pr-147

cccaagcttcggtagacctagaggccttcagcgagtttaccaagatcatcaccccggccatcacccgcgtggtggactttgccaaaaaa ctgcccatgggtacccc

Pr-148

ggggtacccatgggcagttttttggcaaagtccaccacgcgggtgatggccggggtgatgatcttggtaaactcgctgaaggcctctag Gtctaccgaagcttggg

Pr-149

Cccaagcttcgcccatgttctccgagctgccttgtgaagaccagatcatcctcctgaagggctgctgcatggagatcatgtccctgcgg gcagctgtcggtacccc

Pr-150

ggggtaccgacagctgcccgcagggacatgatctccatgcagcagcccttcaggaggatgatctggtcttcacaaggcagctcggagaa catgggcgaagcttggg

Pr-153

cccaagcttcggtcgagtgccccaccgaactcttccccccactcgctctggaggtctttgaggatggtacccc

Pr-154

ggggtaccatcctcaaagacctccagagcgagtggggggaagagttcggtggggcactcgaccgaagcttggg

Pr-155

cccaagcttcggtcgagtgccccaccgaactcttccccacactcttcctggaggtctttgaggatggtacccc

Pr-156

ggggtaccatcctcaaagacctccaggaagagtgtggggaagagttcggtggggcactcgaccgaagcttggg

Pr-159

ggggtaccgagccagaggacactcggt

Pr-160

gctctagactgttcatctttgtccaggca

Pr-161

ggggtaccatggaacagaagcccagcac

Pr-162

gctctagactgttcatctttgtccaggta

Pr-163

cagatggaaagcgaaaaagggcgaacggccaatgtcccc

Pr-164

ggggacattggccgttcgccctttttcgctttccatctg

Pr-165

ggagaacagtgccaggtcagcagctgcaaagcgaaaaaggaagaac

Pr-166

gttcttcctttttcgctttgcagctgctgacctggcactgttctcc

Pr-167

cccaagcttcggcagatggaaagcgaaaaaggaaggcgggtacccc

Pr-168

ggggtacccgccttcctttttcgctttccatctgccgaagcttggg

Pr-169

cccaagcttcggcagatgcaaagcgaaaaaggaaggcgggtacccc

Pr-170

ggggtacccgccttcctttttcgctttgcatctgccgaagcttggg

Pr-171

cccaagcttcggcagctggaaagcgaaaaaggaaggcgggtacccc

Pr-172

ggggtacccgccttcctttttcgctttccagctgccgaagcttggg

Pr-174

cgaactcttccccccactcGCcctggaggtctttgaggatcag

Pr-175

ctgatcctcaaagacctccagggcgagtggggggaagagttcg

Pr-182

cagaactcttcccccctttggccttggaagtgttcgaggattag

Pr-183

ctaatcctcgaacacttccaaggccaaaggggggaagagttctg