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. Author manuscript; available in PMC: 2015 Jul 31.
Published in final edited form as: Cell Rep. 2015 Jul 16;12(4):599–609. doi: 10.1016/j.celrep.2015.06.054

Figure 2. AMPK inhibits GLI1 transcriptional activity.

Figure 2

(A) NIH3T3 cells were treated with 25mM 2-deoxyglucose (2DG), (B) 150 M A769662 and (C) 0.75 mM AICAR for the indicated hours and the amount of Gli1 or Ptch1 mRNA was analyzed by qRT-PCR with Gapdh mRNA as the internal control, and normalized to the time zero Gli1 and Ptch1 mRNA levels. The control is provided by time zero, when no chemicals were applied, so the bars indicate Gli1 and Ptch1 mRNA levels relative to those of Gapdh and normalized to levels at time zero. (D) AMPK+/+ and AMPK -/- MEFs were treated with 0.75 mM AICAR for the indicated hours and was analyzed by qRT-PCR as shown as (A). (E) NIH3T3 cells were serum-starved (SS) in DMEM (0.5% bovine calf serum) overnight, stimulated with Shh with or without 25mM 2DG, for 6 hours, and the amount of Gli1 or Ptch1 RNA was analyzed by qRT-PCR and normalized to Gli1 and Ptch1 mRNA levels in serum-starved cells. Three replicate experiments were done with standard deviations. (F) HEK-293 cells were co-transfected with Gli1-RE-Luciferase reporter, GLI1 and constitutively active AMPK (CA-AMPK), and maintained for 36 hours. Cell lysates were analyzed using a luciferase assay to measure reporter-gene transcriptional regulation by GLI1. Representative results from three experiments (n = 3) conducted in duplicate are shown, with standard deviations.