SIRT1 Attenuates Apoptosis of Nucleus Pulposus Cells by Targeting Interactions between LC3B and Fas under High-Magnitude Compression

Figure 3

SIRT1 overexpression enhanced autophagy and reduced cell apoptosis in rat NPCs under high-magnitude compression. (a) The efficiency of SIRT1 overexpression was detected via quantitative real-time PCR and western blotting. (b) The expression levels of autophagic biomarkers (LC3B and P62) and apoptosis-related proteins (Bcl-2, Bax, and cleaved caspase 3) in each group were analyzed by western blotting. (c) Formation of autophagosomes detected by analyzing SQSTM1/P62 immunofluorescence in NPCs before and after SIRT1 overexpression under high-magnitude compression. (d) Autophagosomes in NPCs were examined via TEM. Red arrows show characteristic double-membrane autophagosome formation. (e, f) The apoptosis rate of the NPCs in each group was analyzed via flow cytometry and TUNEL staining. vs. the 0% (control) compressive deformation group. . . NPCs: nucleus pulposus cells; PCR: polymerase chain reaction.

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