Fig. 2

17β-estradiol increases SCD-1 expression in MCF-7 and T47D cells but not MCF-10A cells. MCF-7, T47D, and MCF-10A cells were incubated for 7 days in phenol red-free media supplemented with charcoal-stripped FBS, followed by a 5 day incubation period in the same media that was supplemented with 2nM 17β-ED or its vehicle EtOH (Ctrl). (a) Cellular proteins were separated by SDS-PAGE and immunoblot analysis of SCD-1 expression was performed using actin as loading control. The graphs show densitometry quantification of the SCD-1 blots. (b) RNA was extracted from cells and reverse transcribed into cDNA. Relative qPCR was performed using HPRT as reference gene for MCF-7 and T47D cells and RNS18S1 for MCF-10A cells. Immunoblots are representative of 3 independent experiments. Data are means ± SEM, n = 3 independent experiments. *Different from control as determined by Student’s t-test (p < 0.05)