Z723975 1ea

After xylol dewaxing, the GBM tumor tissue slices were immersed in hematoxylin solution (C0107, Beyotime, China) for 5 min and then rinsed with distilled water for 1 min. After being differentiated into 75% hydrochloric acid ethanol for 30 seconds, the slices were immersed in 1% eosin solution (C0109, Beyotime, China) for 3 min. After dehydrating, xylene was utilized as transparent. The neutral balsam (D054-1-1, Jiancheng, China) was exploited to seal the slices. In the end, the slices were placed in an optical microscope (Z723975-1EA, Sigma, USA) to observe the tissue integrity.

In this research, the cells in good growth condition were made into a single-cell suspension. Cell suspension (1×10⁴ cells/ml) was seeded into a 6-well plate and then transfected based on the above groups. After incubation for 14 days, we used the 4% paraformaldehyde (P0099, Beyotime) and Giemsa dye solution (G8220, Solarbio, China) to fix and stain cells, respectively. Finally, we used a microscope (Z723975-1EA, Sigma) to count the clone numbers of the cells.

The ileum tissue slices were subjected to dewaxing with xylol, which were then rehydrated with ethanol gradient. After rinsing, we utilized hematoxylin (G1004, Servicebio, China) to stain the slices. After differentiating, the slices were subjected to stain with 1% eosin solution (C0109, Beyotime, China). Then, the results were observed using an optical microscope (Z723975-1EA, Sigma, USA) after dehydrating and sealing. The histopathological score was estimated by two independent pathologists who were unaware of the study conditions based on the histopathological scoring system described by Gao et al. [24 (link)]. Grade 0 represents normal intestine; grade 1 represents separation of the Villous core; grade 2 shows epithelial cell abscission to the level of the middle villi; grade 3 indicates necrosis of the entire villi. Tissues with a histological score of ≥2 are considered to manifest NEC.