Supplementary Figures 1-8 from STAT3-RANTES Autocrine Signaling Is Essential for Tamoxifen Resistance in Human Breast Cancer Cells

Abstract

<p>PDF file - 1.2MB, Supplementary Fig. S1. MCF-7 cells were cultured in the absence or presence of tamoxifen for 9 months. Supplementary Fig. S2. Levels of cytokines secreted by MCF-7 and TRM-7 cells, as determined using a cytokine array. Supplementary Fig. S3. Neutralization of secreted RANTES was confirmed by ELISA. Supplementary Fig. S4. (A) RANTES siRNA constructs evaluated by real-time PCR. (B) STAT3 siRNA constructs evaluated by real-time PCR. The siRNA that resulted in the greatest knockdown of each gene was selected. Columns, means of triplicate samples; bars, S.D. *, P < 0.01. (C) RANTES and total STAT3 mRNA levels in siRNA-transfected MCF-7 and TRM-7 cells were determined by RT-PCR. Supplementary Fig. S5. (A) Recombinant RANTES (50 ng/ml)-treated MCF-7 cells were subjected to western blotting. (B) MCF-7 cells were cultured in the absence or presence of recombinant RANTES (50 ng/ml) for 2 weeks and their sensitivities to actinomycin D and (C) cisplatin were determined. Supplementary Fig. S6. (A), (B) T47D cells were cultured in the absence or presence of recombinant RANTES (50 ng/ml) for 2 weeks. Supplementary Fig. S7. (A), (B) pY-STAT3 and RANTES expression levels in primary ERa-positive breast cancer (n = 8) and tamoxifen treated ERa-positive breast cancer (n = 8) cells were evaluated by immunohistochemical staining. Supplementary Fig. S8. (A) ERa expression levels in MCF-7 and TRM-7 cells. (B) ERK and p38 activation in MCF-7 and TRM-7 cells. (C) RANTES and RANTES receptor (CCR1, 3, 5) mRNA levels were determined by real-time PCR and RT-PCR</p>