Skip to main content

Advertisement

Figure 1 | Molecular Cancer

Figure 1

From: The serine protease inhibitor serpinE2 is a novel target of ERK signaling involved in human colorectal tumorigenesis

Figure 1

SerpinE2 is overexpressed in intestinal epithelial cells transformed by oncogenic MEK1, RAS and BRAF. A- wtMEK- and caMEK-expressing IECs were treated or not with U0126 (20 μM) during 24 h. Thereafter, cells were lysed and RNA isolated for serpinE2 or β-actin gene expression by RT-PCR. B- IEC-6 expressing oncogenic RAS or wtMEK or caMEK were lysed and RNA isolated for serpinE2 or β-actin gene expression by RT-PCR. C- BRAF:ER cells were stimulated with 250 nM 4-hydroxy-tamoxifen (4-OTH) for the indicated time periods. Thereafter, cells were lysed and RNA isolated for serpinE2 or TATA-binding protein (TBP) gene expression by RT-PCR. For the 12 h time point, cells were also treated with 20 μM U0126. ERK1/2 phosphorylation levels and total ERK1/2 were monitored by Western blot. D- wtMEK- and caMEK-expressing cells were treated or not with U0126 (20 μM) during 24 h. Thereafter, equal amounts of concentrated culture medium from wtMEK- and caMEK-expressing cells were analyzed by Western blotting with specific antibodies against serpinE2 as described in Material and Methods. Bovine follicular fluid was used as a positive control (CTL). E- caMEK-expressing IECs were cultured at post-confluence, lysed in Laemmli buffer and analyzed by Western blotting with specific antibodies against serpinE2 (lane 2). The culture medium was concentrated and also analyzed by Western blotting for serpinE2 expression (lane 1). In some experiments, foci were harvested by aspiration with a pipet, lysed in Laemmli buffer (lane 4) and analyzed by Western blotting for the expression of serpinE2. The surrounding cells (monolayer, lane 3) were also lysed in Laemmli buffer for Western blotting with specific antibodies against serpinE2.

Back to article page