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. 2016 May 9;126(6):2308–2320. doi: 10.1172/JCI84465

Figure 3. The miR21/GRHL3 axis regulates D3 expression and TH intracellular concentration.

Figure 3

(A) D3 protein levels were measured by Western blot from BCC cells transfected with miR21, GRHL3, and the 2 expression plasmids together. Lysates were also probed with anti-Flag antibody as control of Flag-GRHL3 transfection efficiency. (B) BCC cells were transfected with miR21 and miR21 plus 2 different siRNAs for GRHL3 knockdown. Anti-GRHL3 antibody was used as internal control of GRHL3 knockdown. (C and D) The Dio3 promoter was cotransfected with the indicated constructs together with CMV-renilla as internal control. Luciferase activity was measured and reported as means ± SD of the LUC/renilla ratios from at least 3 separate experiments, performed in duplicate. (E) The T3-responsive promoter TRE3-TK-Luc was cotransfected in BCC cells with the indicated plasmids. Luciferase activity was measured as in C and D. *P < 0.05, **P < 0.01. (F) The miR21/GRHL3/D3 axis regulates intracellular T3 availability.