Immunofluorescent staining
This protocol is extracted from research article:
RBM5-AS1 promotes radioresistance in medulloblastoma through stabilization of SIRT6 protein
Acta Neuropathol Commun, Jul 5, 2021; DOI: 10.1186/s40478-021-01218-2

DNA damage was assessed by analyzing γ-H2AX foci formation [45]. Briefly, cells were radiated with a dose of 8 Gy, fixed with 4% paraformaldehyde, and permeabilized with 0.2% TritonX-100. After blocking, the cells were incubated with anti-phospho-γ-H2AX (ab26350, Abcam, Cambridge, MA, USA; 1:50 dilution). An Alexa-Fluor 488 conjugated goat anti-rabbit IgG was used as the secondary antibody. Nuclei were counter-stained with 4′,6-diamidino-2-phenylindole (DAPI; Sigma-Aldrich).

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