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. Author manuscript; available in PMC: 2011 May 14.
Published in final edited form as: Mol Cell Endocrinol. 2010 Feb 17;320(1-2):76–86. doi: 10.1016/j.mce.2010.02.014

Figure 4.

Figure 4

GR specific shRNAmir sequences reduce both mRNA and protein expression. 697 and CEM-C7 cells stably transduced with a GR-specific shRNAmir lentivirus were treated with 2-fold serial dilutions of Doxy, starting at a concentration of 1 μg/mL (rightmost sample). The total GR (containing the exon 5/6 junction) transcript concentration was measured with QRT-PCR in 697-shGR2 (A) and C7-shGR1 (B) cells after 48 hours of Doxy pre-treatment followed by 18 hours of Dex treatment. (†) notes, in A, the Dex-induced transcript level after pre-treatment with Doxy that corresponds to the same Doxy concentrations within which the biological response switches (see Fig. 5A ). C & D) Expression levels of GR, turboRFP, and GAPDH proteins were analyzed by western blotting following the same treatment regimen as in A and B. During these studies one 697-shGR2 sample was lost; therefore, in C, the highest Doxy concentration analyzed was 500 ng/mL. This does not affect the interpretation of the data.