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. 2009 Jan 1;23(1):24–36. doi: 10.1101/gad.1753809

Figure 6.

Figure 6.

TGF-β and activated Kras signaling impact Gli and Ptch1 expression in a Smo-independent manner. (A) Expression of Smo, Ptch1, E-Cad, Gli1, and Gli3 mRNA in total RNA extracts from wild-type Smo (4.2 NR) or Smo mutant (4.2 R) PDAC cell lines 48 h after stimulation with 5 ng/mL recombinant TGF-β1. Levels of mRNAs expressed as a percentage of the mGus control mRNA. (B) Expression of Kras, Gli1, and Ptch1 mRNA in total RNA extracts from wild-type Smo (4.2 NR) or Smo mutant (4.2 R) PDAC cell 48 h after transfection with control siRNA pools or siRNA pools targeting Kras or Gli1. Levels of mRNAs expressed as a percentage of the mGus control mRNA. (C) Absorbance at 540 nm of two PDAC cell lines (4.2 NR and 3.3) incubated with MTT (see the Materials and Methods) 72 h after transfection with control siRNA pools or siRNA pools targeting Kras or Gli1 and after 24 h of serum starvation. (D) Relative change in Activated Caspase 3 immuno-fluorescent staining of mouse PDAC 3.3 cells 60 h after transfection with control siRNA pools (Ctrl) or siRNA pools targeting Kras or Gli1, following 12 h of serum starvation. Staining was evaluated in five fields each containing at least 500 cells for each condition. Act-Casp-3-positive cells expressed as a percentage of DAPI nuclei; the percentage in Ctrl-treated cells was set at 100%. Asterisk indicates a P-value <0.01 (A), <0.05 (B,C), or <0.005 (D).