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. Author manuscript; available in PMC: 2012 Sep 1.
Published in final edited form as: Cancer Res. 2011 Jul 11;71(17):5904–5914. doi: 10.1158/0008-5472.CAN-10-4173

Figure 4.

Figure 4

Gli3R and GANT61 produce similar cellular responses in HT29 cells. A: Transient transfection with Gli3R for 72 hr, or B: Treatment with GANT61 (20 µM) for 72 hr. Untransfected (UT), empty vector transfected (Vector), GLI3R transfected (Gli3R) cells, or untreated (UT), DMSO- or GANT61-treated cells, were examined under an inverted microscope at 72 hr to determine changes in cellular morphology (left panels). Cells were also counted using a coulter counter to monitor cell growth per well. Cell death was determined at 72 hr by flow cytometry following Annexin V-FITC/PI staining. Total cell lysates were extracted from treated cells at 0 hr, 24 hr, 48 hr and 72 hr; 60 µg of total protein was analyzed by western blot using antibodies specific for the myc tag, Gli1, Gli2 and HSP90α/β as the loading control. C: Expression of γH2AX, cleaved PARP and cleaved caspase-3 determined by western analysis.