Fig. 6

The induction of SCD-1 by 17β-estradiol partially involves IGF-1R. MCF-7 cells were incubated for 7 days in phenol red-free media supplemented with charcoal-stripped FBS (starved), followed by an incubation in the same media that was supplemented with 50 ng/ml of IGF-1, 2nM of 17β-ED, 10 μM of the IGF-1R antagonist Ag1024, a combination of 17β-ED, and Ag1024, or their vehicle controls (Ctrl) for 5 days. (a) Cells subjected to the different treatments were counted using a haemocytometer. (b) Cellular proteins were separated by SDS-PAGE and immunoblot analysis of SCD-1 levels was performed using actin as loading control. The graphs show densitometry quantification of the SCD-1 blots. (c) RNA was extracted from cells and reverse transcribed into cDNA. Relative qPCR for SCD-1 was performed, using HPRT as reference gene. Immunoblots are representative of 3 independent experiments. Data are means ± SEM, n = 3 or 4 independent experiments. Values that have a different superscript are significantly different (p < 0.05) as determined by one-way ANOVA test with subsequent Tukey’s adjustment