Fig. 6

AR acts as a direct regulator of Ucp1 expression in the 3T3-L1 cells in vitro. A adipogenic differentiation of 3T3-L1 cultured in undifferentiated condition (naïve), differentiated with normal serum (FBS) and differentiated with steroid-depleted serum (csFBS). Upper images show phase contrast of three experimental groups. Lower images show neutral lipid staining with ORO. Images were taken at 200X of magnification. B In Ucp1 expression levels under steroid-depleted FBS for 5,10 and 15 days (upper panel). Ucp1 expression under steroid-depleted FBS, androgen replacement by 5 nM DHT and specific blockade of AR by 20 µM bicalutamide (CDX, lower panel). C Ucp1 expression after incubation for 24 h with CDX and withdraw for 24 h (left) and 72 h (right). D 2% agarose gel of PCR products derived from ChIP. (IS) input sample, (IP) immunoprecipitated sample, (DMSO) vehicle, (CDX) bicalutamide. E Sequence alignment of ChIP results vs complete Ucp1 sequence. Scheme represents which nucleotide correspond to every position. (A) adenine, (C) cytosine, (G) guanin, (T) thymine, (N) non detectable. Frequency represents the conservation of this position between sequences. Symbol height represents the relative frequency for this nucleotide in the predicted position. F DNA fold enrichment vs input after immunoprecipitation in 3T3-L1 fibroblast incubated with vehicle (DMSO) or 20 µM bicalutamide (CDX). n = 3. Data were represented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001