Ab267389

Formalin-fixed, paraffin-embedded LUAD and adjacent normal tissues were fixed by 10% formalin for 48 h. The tissues were sectioned into 4 μm thickness. Afterwards, the slices were deparaffinized utilizing xylene as well as antigen retrieval. After blockage, the sections were incubated with primary antibody against HOXA1 (1 : 100; ab72591; Abcam, USA), Nrf2 (1 : 100; 16396-1-AP; Proteintech, China), HO-1 (1 : 100; 27282-1-AP; Proteintech, China), and CD155 (1 : 100; ab267389; Abcam, USA) overnight at 4°C, followed by HRP-labeled secondary antibodies (1 : 200; ab97080; Abcam, USA) for 30 min at room temperature. After being stained by hematoxylin, the sections were scanned with a PathScope pathology slide scanner.

Total proteins were extracted from tissue or cell specimens utilizing RIPA lysis, which were determined with BCA kit. 20 μg proteins was separated via 10% SDS-PAGE and transferred onto PVDF membranes. After being blocked, they were incubated at 4°C for 12 h with primary antibodies against HOXA1 (1 : 1000; ab72591; Abcam, USA), Nrf2 (1 : 1000; 16396-1-AP; Proteintech, China), HO-1 (1 : 1000; 27282-1-AP; Proteintech, China), CD155 (1 : 1000; ab267389; Abcam, USA), and β-actin (1 : 5000; ab179467; Abcam, USA), followed by incubation with secondary antibodies (1 : 5000; ab7090 or ab7097; Abcam, USA) at room temperature for 2 h. Protein band was visualized with ECL kit. Protein expression was quantified utilizing ImageJ software, with β-actin as the loading control.

Cervical biopsy samples were routinely embedded in paraffin and sectioned at a thickness of 3 μm, followed by hematoxylin–eosin (HE) staining. The HE-stained tissue sections were then examined by three experienced pathologists. Prior to IHC staining, the tissue sections were dewaxed in xylene and rehydrated in graded alcohol. The tissue sections were incubated with the following antibodies: PVR (ab267389, diluted at a ratio of 1:300, Abcam), E-Cadherin and N-Cadherin (MAB0738 and MAB0571, respectively, diluted at a ratio of 1:200, MXB Biotechnologies), followed by IHC staining using the streptavidin-peroxidase conjugate method. Negative controls were prepared with non-immune isotype-specific sera used in place of primary antibodies, while previously positively-stained tissue sections for the labeled proteins were taken as positive controls.