Figure 8

Importance of individual binding site for Dcx promoter activity. (a-b-c) ESR1 cells were transfected with the indicated constructs at day of differentiation 0, DD0 (a), DD6 (b) on gelatin and at DD18 (c) on poly-ornithine/laminin and luciferase activity relative to β-galactosidase activity was determined 48 hours later. The different constructs were pdcx 249bp-Luc, 79bp-deleted pdcx 249bp-Luc construct (pdcx 249bp^--Luc) and pdcx 249bp-Luc with mutated binding site (pdcx 249bp/Lef*, pdcx 249bp/Hnf6*, pdcx 249bp/Nf-y2* and pdcx 249bp/Nf-y3*) and the promoterless control (CTL). (d) CGN were isolated at post-natal day 3 and were transfected the same day with the indicated constructs and luciferase activity relative to β-galactosidase activity was determined 48 hours later. The different constructs were pdcx 249bp-Luc, 79bp-deleted pdcx 249bp-Luc construct (pdcx 249bp^--Luc) and pdcx 249bp-Luc with mutated binding site (pdcx 249bp/Lef*, pdcx 249bp/Hnf6*, pdcx 249bp/Nf-y2* and pdcx 249bp/Nf-y3*) and the promoterless control (CTL). The activity of each construct is expressed relative to that of the pdcx 249bp-Luc construct, set arbitrarily to 1. Note that the relative activity of pdcx 249bp-Luc, used here as a reference was consistent with the results shown in figure 2. Each value represents the mean ± SEM of at least three independent transfection experiments, each performed in triplicate. Asterisks mean significantly different from pdcx 249bp values at P < 0.05 (*) or P < 0.01 (**).