Figure 4. RT-PCR and Western blot verification of microarray data for selected pancreatic cancer cell lines.

(A) RT-PCR was performed on lysates from the indicated cell lines, and intensities of the bands at the expected size for each gene were quantified. A student's t-test (results given by the p value) was performed comparing the band intensities between the mesenchymal-like cell lines (in bold) and the epithelial cell lines. In addition, a correlation was calculated (results given by the r value) between the RT-PCR results and the microarray data. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as a cDNA loading control. (B) Comparison of GALNT3 protein levels by Western blot in selected pancreatic cancer cell lines. Lysates were collected from the selected cell lines, fractionated by SDS-PAGE, and probed by Western blot. Mesenchymal-like cell are labeled in bold. The highlighted bands are at the expected molecular weights of 73 kD for GALNT3 and 42 kD for actin. (C) MRC2 protein levels measured by Western blot in TGFβ treated and untreated cells. The cell lines were either exposed to 5 ng/mL TGFβ or untreated under serum starvation for 72 hours prior to cell lysis. The highlighted bands are at the expected molecular weights of ∼167 kD for MRC2 and 42 kD for actin.