Paraffin embedded zinc-formalin fixed tissue was sectioned and antigen retrieval was performed with citrate pH6.0 buffer
in a pressure cooker. Slides were blocked with serum-free protein block (X0909, Dako) for 1 hour at RT and incubated overnight at
4°C with the following antibodies FOXM1 (AF3975, Novus, 1:100), PRRX1 (NBP2-13816, Novus, 1:50) or GFP (ab13970, abcam,
1:250). Immunohistochemistry was performed using Biotinylated secondary antibodies, ABC kit (PK-6100) and DAB (SK-4100) from
Vector Labs. For immunofluorescence, slides were incubated with Cy-conjugated secondary antibodies (Jackson ImmunoResearch, 1:300)
for 1 hour at RT and nuclei were stained with DAPI. Image were acquired on a Nikon E600 microscope and processed with ImageJ
(1.51w) software. Antibody fidelity was verified by staining murine PDAC tissue in the absence of primary antibody (Supplementary Figure 1A ). Human PDAC tissue was obtained from US Biomax,
Inc. (PA483).
in a pressure cooker. Slides were blocked with serum-free protein block (X0909, Dako) for 1 hour at RT and incubated overnight at
4°C with the following antibodies FOXM1 (AF3975, Novus, 1:100), PRRX1 (NBP2-13816, Novus, 1:50) or GFP (ab13970, abcam,
1:250). Immunohistochemistry was performed using Biotinylated secondary antibodies, ABC kit (PK-6100) and DAB (SK-4100) from
Vector Labs. For immunofluorescence, slides were incubated with Cy-conjugated secondary antibodies (Jackson ImmunoResearch, 1:300)
for 1 hour at RT and nuclei were stained with DAPI. Image were acquired on a Nikon E600 microscope and processed with ImageJ
(1.51w) software. Antibody fidelity was verified by staining murine PDAC tissue in the absence of primary antibody (
Inc. (PA483).