Ab207718

The following antibodies were purchased from the respective suppliers: anti-RNPS1 (ab79233), anti-Ki-67 (ab15580), anti-CEA (ab207718), anti-CA199 (ab3982), anti-CA153 (ab109185), anti-HE4 (ab200828), anti-Bcl-2 (ab182858), anti-Bax (ab182733), anti-MLH1 (ab92312), anti-cleaved caspase-3 (ab2302), anti-MSH2 (ab212188), anti-MSH6 (ab92471), and anti-PMS2 (ab110638) (all from Abcam, Cambridge, USA) and anti-β-actin (M01263-2; Boster, Wuhan, China). The secondary antibodies used were anti-rabbit IgG (AS014) and anti-mouse IgG (H+L) (AS003), both from ABclonal (Wuhan, China), and IMR-1A (HY-100431A; MedChemExpress, Dallas, TX, USA).

Gastric paracarcinoma and cancerous tissues from animal models were fixed with 4% paraformaldehyde for 48 h, dehydrated, and embedded in paraffin. The paraffin sections were subjected to antigen retrieval, sealing, incubation with primary antibodies (1:50 dilutions of myeloperoxidase [MPO] [ab90810; Abcam, UK], cit-H3 [ab5103; Abcam], CD66b [ab207718; Abcam], CCDC25 [21209–1-AP; Proteintech, China], and CD31 [ab228968; Abcam]), incubation with secondary antibodies (1:100 dilutions of horseradish peroxidase [HRP]-labeled goat anti-mouse IgG and HRP-labeled goat anti-rabbit IgG), 4′,6-diamidino-2-phenylindole (DAPI) staining (Solarbio Life Science, Beijing, China), coloration, and sealing. The samples were then observed and photographed using a confocal microscope (LSM800; Zeiss, Germany).

We collected pathologically confirmed pancreatic cancer specimens from 20 patients undergoing radical pancreatic cancer surgery from the Peking Union Medical College Hospital (PUMCH) with the approval of the Institutional Ethics Committee of PUMCH (Ethic code: I-22PJ487). Twenty surgical pancreatic cancer specimens were fixed in formalin and then embedded in paraffin. Multispectral IF staining was performed as previously described^{62 (link)}. These tumor tissue sections were incubated with the following antibodies: anti-CD66b (Abcam, ab207718, dilute at 1:500), anti-Myeloperoxidase (Abcam, ab208670, dilute at 1:100), anti-CD68 (Abcam, ab213363, dilute at 1:100), and anti-LC3B (Abcam, ab192890, dilute to 1 µg/ml). Nuclear staining was performed with ProLong Diamond Antifade mounting medium containing DAPI (Invitrogen). Images of tissue specimens were obtained using the TissueFAXS Spectra Systems (TissueGnostics GmbH, Vienna Austria). With the help of StrataQuest analysis software (Version 7.1.129, TissueGnostics GmbH, Vienna, Austria), we separated the multi spectral image (5-color staining) and set a threshold to divide the positive cells for analysis of cell number and location distribution.