Hypoxia Helps Maintain Nucleus Pulposus Homeostasis by Balancing Autophagy and Apoptosis

Figure 3

Autophagosome- and autophagy-related genes and protein level analysis in NP cells under different oxygen concentrations. (a) Representative MDC staining and TEM images and statistical analysis of autophagosomes (arrowheads) in p5, p15, and p20 NP cells. Scale bar for MDC staining: 100 μm, TEM: 0.5 μm. (b) RT-PCR was used to analyze mRNA expression levels in normoxic (C-NP) and hypoxic (H-NP) cells at p5, p15, and p20 ( per group). Primers targeted hypoxia- (HIF1, SIRT1) and autophagy- (HMGB1, beclin-1, ATG7, ATG5, LC3-I, and LC3-II) related genes. (c) Representative immunoblots and statistical analysis of protein levels in the experimental groups. Data are presented as the . , , and versus p5; ^#, ^##, and ^### versus p15; ^†, ^††, and ^††† versus C-NP cells at the same passage number.

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