Figure 5
Effects of asPR or RU 486 on MAPK phosphorylation and on ER-α and PR expression. (a) Immunoblots of ER-α (MC-20; Santa Cruz Biotechnology), total ERK (K-23; Santa Cruz Biotechnology), and pERK (E-4; Santa Cruz Biotechnology) in whole extracts of tumors obtained from animals treated with saline, asPR, or scPR for 5 days. Tumor samples were obtained after day 5 of treatment; tumor growth kinetics is shown in Fig. 2d. Arrows show ERK1 (42 kDa) and ERK2 (44 kDa). PR immunoblots were performed using extracts obtained from mice treated with asPR over 10 days (Ab1; Dr Shyamala). Arrows show the classical PRB of 115 kDa and the classical PRA of 83 kDa. (b) Immunoblots of ER-α, PR (Ab7; Neomarkers), E-cadherin (E cad; BD Transduction Lab), total ERK, and pERK using wholeextracts of tumors obtained from animals treated with saline or RU 486 for 24 hours. Tumor kinetics are shown in Fig. 2c. A representative Western blot of three is shown. (c) Immunohistochemistry of ER-α and PR (C-20 Santa Cruz) of the same tumor samples used in Western blot studies shown in panel b (125×). Experimental details are described in Materials and method. asPR, antisense oligodeoxynucleotides to progesterone receptors; ER, estrogen receptor; ERK, extracellular signal-regulated kinase; MAPK, mitogen-activated protein kinase; PR, progesterone receptor; scPR, scrambled oligodeoxynucleotides to progesterone receptors.