Nbp1 32440

Fallopian tube and high-grade ovarian carcinoma sections were processed as previously reported (55 (link)). The immunohistochemical staining were performed using a dilution of 1:500 of antibodies to PAX8 (Novus: NBP1-32440) or SOX17 (Novus: NBP1-47996). Slides were scanned with Aperio CS2.

10⁴ cells from each FTSEC and HGSOC line were seeded onto imaging plates (Eppendorf: 0030741030) and allowed to grow for 24 hours. Cells were washed twice in PBS, and fixation was performed for 15 min with paraformaldehyde 4% (Thermo-Fisher: AAJ19943K2) at room temperature. Cells were then washed twice in PBS and permeabilized with Triton X-100 0.25% (Boston BioProducts: P-924) for 15 min. Aldehyde residues were quenched with glycine 100 mM (Sigma-Aldrich: 50046-50G) for 15 min. The unspecific sites were blocked with a solution of 1% bovine serum albumin, and 0.1% Tween 20 in PBS for 30 min. Samples were incubated for 16 hours at 4 °C with a dilution of 1:500 of antibody to PAX8 (Novus: NBP1-32440) or SOX17 (Novus: NBP1-47996). Cells were then washed three times for 5 min each in a solution of 1% bovine serum albumin and 0.1% Tween 20 in PBS followed by incubation for one hour with AlexaFluor488-conjugated anti-mouse IgG antibody or AlexaFluor594-conjugated anti-rabbit IgG antibody. Cells were washed three times in PBS, mounted in Fluoromount-G with DAPI, and images were acquired at 60X magnification with a Nikon Eclipse Ti inverted microscope.