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. 2014 Sep 8;42(17):11011–11024. doi: 10.1093/nar/gku814

Figure 4.

Figure 4.

Plau-001 and Plau-004 mRNAs in MDA-MB231 cells. (A) Structure of the Plau locus: Plau-001 is depicted as in Figure 2A. Plau-004 is depicted as described in the ENSEMBL.org database. The dashed line and the question mark indicate that its 3′ end has not been characterized. (B) Plau-004 mRNA expression in MDA-MB231 cells. The sketch represents Plau-004 exon 1 and 2. Amplicon 1 and 2 correspond to non-spliced RNA, whereas amplicon 3 corresponds to spliced RNA. RT-PCR products from total MDA-MB231 cells were analyzed by agarose gel electrophoresis. (C) Relative abundances of Plau-001 and Plau-004 in MDA-MB231 cells. RNA abundances were assayed by RT-qPCR from total cellular RNA. RNA levels were normalized to that for S26 and the value for Plau-004/S26 was set to 1. (D) Plau-004 contains Plau-001 3′UTR. Reverse transcription was performed using total RNA from MDA-MB231 cells and random primers. PCRs were then carried out using a 5′ primer located in Plau-004 exon 1 and 3 downstream primers located at different positions in the uPA locus as depicted in the figure. PCR products were analyzed by agarose gel electrophoresis. (E) Expression of Plau-004 in different aggressive breast cancer cell lines. Total RNA was prepared from various breast cancer cell lines, which all express substantial amounts of Fra-1 and uPA mRNA except MCF-7. RT-PCR products corresponding to amplicon 3, as described in (B), were analyzed by agarose gel electrophoresis.