A Cy3-labeled SLC7A11-AS1 probe was generated by RiboBio (Guangzhou, China). Slides were xed with 4% paraformaldehyde for 10 min at room temperature, followed by permeabilization at 4°C for 5 min. The slides were hybridized overnight with the SLC7A11-AS1 probe and then washed three times with a graded saline sodium citrate solution.
For immuno uorescence staining, cells were xed with 4% paraformaldehyde for 30 min, stained with a rabbit anti-xCT antibody (1:100 dilution; Proteintech, China) overnight, washed with PBS three times, incubated with a Cy3-goat anti-rabbit IgG (H+L) secondary antibody (1:50 dilution; Beyotime, China) and washed with PBS three times. The nuclei were counterstained with DAPI. Fluorescence images were acquired with a confocal microscope (Leica, Wetzlar, Germany).
For immuno uorescence staining, cells were xed with 4% paraformaldehyde for 30 min, stained with a rabbit anti-xCT antibody (1:100 dilution; Proteintech, China) overnight, washed with PBS three times, incubated with a Cy3-goat anti-rabbit IgG (H+L) secondary antibody (1:50 dilution; Beyotime, China) and washed with PBS three times. The nuclei were counterstained with DAPI. Fluorescence images were acquired with a confocal microscope (Leica, Wetzlar, Germany).