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. 2014 May 19;9(5):e96497. doi: 10.1371/journal.pone.0096497

Figure 5. Only GR protein is detected in primary cervical epithelial cells (VEN-100).

Figure 5

(A) Upon arrival the VEN-100 cells were rested overnight before being washed once with PBS and harvested with TRIzol®. Total RNA was isolated and 500 ng RNA was reverse-transcribed. Steroid receptor gene expression was measured by qRT-PCR with receptor-specific primers, followed by gel electrophoresis to confirm the PCR products. (B) VEN-100 cells were rested overnight before harvesting in 2X SDS sample buffer. COS-1 cells were transiently transfected with 1 µg/well pcDNA3 (empty vector) which served as negative control (−CTRL) or with 1 µg/well steroid receptor expression vectors (GR, PR-B, AR, MR and ERα) which served as positive controls (+CTRL). Twenty fourhrs later, the COS-1 cells were washed once and lysed with 2X SDS sample buffer. Equal volumes of cell lysate (VEN-100 and COS-1 ctrls) were analysed by Western blotting with antibodies specific for the GR and Flotillin-1 (loading control), respectively.