Image m1

Paraffin-embedded tumor tissues were sectioned to a thickness of 5 μm,
deparaffinized with xylene, rehydrated through xylene and graded alcohol and
blocked with 5% bovine serum albumin. Immunofluorescence (IF) staining was
conducted with the indicated antibodies and fluorochrome-conjugated secondary
antibodies (Alexa-488 and 564). Apoptotic cells were identified by terminal
deoxynucleotidyl transfer-mediated dUTP nick-end labeling (TUNEL) staining using
an in situ BrdU-Red DNA fragmentation assay kit (Abcam,
Cambridge, UK). Nuclei were counterstained with DAPI. The slides were examined
in a blinded manner and randomly chosen fields were photographed at 400x
magnification. The immune-positive cells were quantified with a Carl Zeiss
AxioImager microscope and Image M1 Software (Carl Zeiss, Jena, Germany).

The fixed tissues were embedded into paraffin blocks, and 5 μm sections were prepared. The tissue sections were picked up onto a glass slide and deparaffinized, rehydrated, and subjected to Hematoxylin and Eosin (H&E) (Merck, Darmstadt, Germany) staining. A Carl Zeiss AxioImager microscope and Image M1 Software (Carl Zeiss, Jena, Germany) were used to provide images of randomly chosen fields at 400x magnification.

Tumor and lung tissues were fixed in formalin (10%) and then parafinized, trimmed and sliced into 5 µm slices. Following mounting the slice on a glass slide, tissue slides were deparaffinized, rehydrated and subjected to H&E staining. Selected fields were randomly photographed at 400× magnification using a Carl Zeiss AxioImager microscope and Image M1 Software (Carl Zeiss, Jena, Germany).