Supplementary Figure 1.
NE fails to activate GR in TNBC cell lines. A. RNA was isolated from a panel of breast
cancer cell lines and adrenergic receptors were measured by real time PCR. ADRB2
expression was not significantly elevated in TNBC cell lines comparative to cell lines
expressing ER+, PR+ Her2+. A. GR activation in MDA-MB-231 cells B. and HCC1187 cells
by luciferase C. GR expression was significantly increased by dexamethasone but not by NE
in each cell line.
Supplementary Figure 2.
Stress hormones induce DNA damage in MCF-7 cells. MCF-7 cells were incubated with
Cort or NE for 24 h and assessed for DNA damage by the comet assay (n = 3). Both Cort and
NE induced significant DNA damage in MCF-7 cells.
Supplementary Figure 3.
Stress hormones induce phospho ATR, p21, and phospho H2AX in a dose dependent
manner. Cell lysates were prepared and resolved by SDS page and analyzed for each protein
target by western blotting. Theoretical molecular masses in kilodaltons are shown. 10 -6 M
cortisol and 10-7 M NE resulted in the most robust changes in protein expression.
Supplementary Figure 4.
Stress hormones induce p. Cell lysates were prepared and resolved by SDS page and
analyzed for p21 by western blotting. Cells were incubated with Cort or NE in the presence
or absence of receptor antagonists, RU-486 and propranolol for 2, 6 and 24 hours and cell
lysates were prepared and resolved by SDS page and analyzed for p21 by western blotting.
Theoretical molecular masses in kilodaltons are shown.
Supplementary Figure 6.
HCC1187 cells were incubated with Cort or NE in the presence or absence of receptor
antagonists, RU-486 and propranolol for 2 and 6 hours and cell lysates were prepared and
resolved by SDS page and analyzed for phospho ATR (ser 428) and phospho Chk1 (ser 345)
by western blotting. Stress hormones exhibited similar effects in HCC1187 cells and induce
ATR, CHK-1 and P21 in the HCC1187 cell line. Note the increased basal expression of ATR.
Theoretical molecular masses in kilodaltons are shown.