Autophagic Degradation of Gasdermin D Protects against Nucleus Pulposus Cell Pyroptosis and Retards Intervertebral Disc Degeneration In Vivo

Figure 3

Detection of autophagy activity in NP cells in vitro. (a) The mRNA level of ATG5 in ATG5 siRNA-treated NP cells was analyzed by qRT-PCR. si-ATG5-1 was used in the following experiments. vs. the control group; ns, , no significant difference. (b) Western blot analysis of LC3, Beclin-1, and P62. LPS-treated (1 μg/mL, 24 h) NP cells were cotreated with 3-MA (10 mM, 24 h) or rapamycin (1 μM, 24 h). Quantification of (c) LC3-II, (d) Beclin-1, and (e) P62 protein levels. GAPDH was used as an internal control. (f) Immunofluorescence analysis showed the expression level of LC3 and (g) relative mean fluorescence was calculated. Data were presented as the , . and vs. the LPS group; ns, , no significant difference. (h) TEM images of NP cells indicated the number and morphology of autophagosomes (red arrowheads).

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