| Id |
Subject |
Object |
Predicate |
Lexical cue |
| T96 |
0-95 |
Sentence |
denotes |
Figure 3 Analysis of the Minimal Region (36 bp) of the Proximal ɛy Promoter Responsive to Sox6 |
| T97 |
96-172 |
Sentence |
denotes |
(A) The sequence of the 36-bp fragment and its mutant versions used in EMSA. |
| T98 |
173-318 |
Sentence |
denotes |
The WT 36-bp DNA sequence (−63 to −28) of the ɛy globin proximal promoter contains two Sox/Sox6 consensus binding sites, shown in bold underline. |
| T99 |
319-447 |
Sentence |
denotes |
Versions with truncation of this sequence (M1) or mutation of one of the two consensus binding sites (M2 and M3) are also shown. |
| T100 |
448-480 |
Sentence |
denotes |
(B) EMSA with c-Myc-tagged Sox6. |
| T101 |
481-653 |
Sentence |
denotes |
EMSA was performed using the 36-bp radio-labeled WT probe (as shown in (A)) and c-Myc tagged Sox6 translated in vitro using reticulocyte lysate (see Materials and Methods). |
| T102 |
654-1039 |
Sentence |
denotes |
Lane 1: radio-labeled free probe (run out of the gel); Lane 2: no competition, no antibody; Lane 3: competition with 200-fold excess cold probe, no antibody; Lane 4: no competition, c-Myc antibody (producing a supershift); Lane 5: no competition, Sox6 antibody (producing a supershift); Lane 6: no competion, no antibody using in vitro translated vector containing c-Myc, but not Sox6. |
| T103 |
1040-1069 |
Sentence |
denotes |
(C) EMSA with HA-tagged Sox6. |
| T104 |
1070-1150 |
Sentence |
denotes |
EMSA was performed similarly as in (B) using HA-tagged Sox6 translated in vitro. |
| T105 |
1151-1370 |
Sentence |
denotes |
Lane 1: radio-labeled free probe (run out of the gel); Lane 2: no competition, no antibody; Lane 3: competition with 200-fold excess cold probe, no antibody; Lane 4: no competition, HA antibody (producing a supershift). |
| T106 |
1371-1435 |
Sentence |
denotes |
(D) EMSA using MEL cell nuclear extracts and the 36-bp WT probe. |
| T107 |
1436-1657 |
Sentence |
denotes |
Lane 1: radio-labeled free probe (run out of the gel); Lane 2: no competition, no antibody; Lane 3: competition with 200-fold excess cold probe, no antibody; Lane 4: no competition, Sox6 antibody (producing a supershift). |
| T108 |
1658-1751 |
Sentence |
denotes |
(E) EMSA with c-Myc-tagged Sox6, WT and mutant versions of the 36-bp fragment in competition. |
| T109 |
1752-1856 |
Sentence |
denotes |
EMSA was performed using the radio-labeled 36-bp WT probe and the c-Myc tagged Sox6 translated in vitro. |
| T110 |
1857-1927 |
Sentence |
denotes |
Lane 1: radio-labeled free probe; Lane 2: no competition, no antibody. |
| T111 |
1928-2060 |
Sentence |
denotes |
Competition was performed using 200-fold excess cold probes corresponding to WT (Lane 3), M1 (Lane 4), M2 (Lane 5), and M3 (Lane 6). |
| T112 |
2061-2140 |
Sentence |
denotes |
(F) Both consensus Sox/Sox6 binding sites are required for Sox6 responsiveness. |
| T113 |
2141-2307 |
Sentence |
denotes |
GM979 cells were transfected with a reporter construct (Figure 2A) containing −63 to +45 of the ɛy proximal promoter together with the CMV-Sox6 overexpression vector. |
| T114 |
2308-2396 |
Sentence |
denotes |
Mutations of the consensus binding sites were also tested (M3, M2, M2 plus M3, see (A)). |
| T115 |
2397-2467 |
Sentence |
denotes |
The fold repression of Sox6 with the WT or mutant constructs is shown. |
| T116 |
2468-2566 |
Sentence |
denotes |
The baseline activities of the mutagenized reporter constructs are comparable to the WT construct. |
