Fig. 1

(A-D). IGF-1 mediates FASN expression in breast cancer through mTORC1. A MDA-MB-231 and MCF-7 cells were serum starved 16 hours followed by either 0, 50, or 100 ng/mL of IGF-1 exposure for 6 hours in full growth media prior to RT-qPCR analysis. Data are represented as a mean of 3 independent trials ± SD. Values are expressed as a relative fold change to no IGF-1 exposure and were standardized to actin. Western blot analysis below the graphs shows an increase in FASN protein expression following IGF-1 exposure (100 ng/mL) for 24 hours. GAPDH was used as a loading control. Band quantification of the 3 trials (n = 3) is expressed as a fold change to no IGF-1 treatment. GAPDH was used to standardize the protein band intensity. All blots in Fig. 1 were cropped to simplify data interpretation. Full blots can be found in Fig. S1. B MDA-MB-231 and MCF-7 cells were untreated or transfected with either a control siRNA or IGF-1R siRNA for 48 hours. Cells were then exposed with or without IGF-1 (100 ng/mL) for 24 hours in full growth media prior to western blot analysis. Band quantification is represented as a graph below expressing the percent change of IGF-1R and FASN relative to the GAPDH loading control. Bars are represented as a mean of three band intensities from 3 independent trials (n = 3) ± SD. C MDA-MB-231 and MCF-7 cells were serum starved 16 hours with vehicle or rapamycin (100 nM) followed by IGF-1 exposure (100 ng/mL) for 6 hours in full growth media prior to RT-qPCR analysis. Graphs are expressed as a fold change to vehicle and represent an average of three independent trials ± SD. All values were standardized to actin. D MDA-MB-231 and MCF-7 cells were serum starved with vehicle or rapamycin (100 nM) and exposed to IGF-1 (100 ng/mL) for 24 hours before western blot analysis. Bar graphs to the right represent an average band quantification from three trials (n = 3) ± SD. Bars represent a fold change to DMSO treatment. p-S6K band intensity was standardized to total S6k and FASN standardized to GAPDH. All statistical analyses in A-D were performed by students T test