Anti src 1

Cells were lysed in RIPA buffer (25mM Tris-HCl pH 7.6, 150mM NaCl,
1% sodium deoxycholate, 0.1% SDS) with Halt TM Protease
Inhibitor Cocktail (Thermo Fisher, Florence, KY, USA) on ice for 30 mis. Samples
were centrifuged at 4 °C, 13,000 g for 15 min. Collected supernatants
and protein concentrations of the supernatants were quantified using the Protein
Quantitation Kit (Abcam, Cambridge, MA, USA). Equal levels of total protein from
different samples were subjected to SDS-PAGE analysis and signals were detected
using corresponding primary and secondary antibodies. The following antibodies
(1:1000 dilution) were used: anti-CXCR4 (Abcam, #ab181020) anti-SRC-1
(Cell Signaling Technology, 128E7, #2191); anti-Glucocorticoid Receptor
(Cell Signaling Technology, D8H2, #3660); anti-p300 (Cell Signaling
Technology, C-20, sc-585); anti-actin (Sigma, A3853).

Cells were lysed in immunoprecipitation (IP) buffer (50 mM Tris-HCl pH
7.5, 150 mM NaCl, 1 mM EDTA, 1% Triton X-100 and protease inhibitor
cocktail) on ice for 30 min. Lysates were centrifuged at 4 °C, 13,000 g
for 15 min and the supernatant was subjected to immunoprecipitation using
primary antibodies or IgG control and protein A agarose beads (Cell Signaling
Technology, Beverly, MA, USA). Precipitated immunocomplexes were washed five
times with washing buffer (50 mM Tris-HCl pH 7.5, 150 mM NaCl, 1 mM EDTA, and
1% Triton X-100) and subjected to immunoblotting analysis. The following
antibodies were used: anti-SRC-1 (Cell Signaling Technology, 128E7,
#2191); anti-Glucocorticoid Receptor (Cell Signaling Technology, D8H2,
#3660); anti-p300 (Cell Signaling Technology, C-20, sc-585); anti-actin
(Sigma, A3853).