Research Paper Volume 15, Issue 24 pp 15382—15401

Proteomic analysis reveals the aging-related pathways contribute to pulmonary fibrogenesis

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Figure 5. Key molecular players of ferroptosis and autophagy signaling pathways in aging. (A) Flow cytometry assay showed that ROS expression increased in senescent cells compared with normal ones. (B) Senescent cells expressed more ferric ions than normal ones. (C) Total iron increased in lung tissue of 16-month-old mice compared with the 2-month-old group. (D) MDA increased in lung tissues of 16-month-old mice compared with the 2-month-old group. (E) Western blot showed that GPX4 expression decreased in aging cells compared with normal cells in vivo and in vitro. (F) qRT-PCR results showed that lncIAPF expression increased with age in a group of healthy population. (G) qRT-PCR results showed that lncIAPF expression increased in the aging cell model than those in the normal one. (H) RIP results verified that HuR specifically concentrated large quantities of lncIAPF. (I) Western blot confirmed that the expression levels of ATF3 and HuR increased in the aging cell model and lung tissue compared with those in the normal group. (J) Application of dual-fluorescence mRFP-GFP-LC3 adenovirus monitoring to observe autophagy under laser confocal microscopy revealed that the number of yellow autophagosomes increased in the aging group compared with the normal one. (K) Western blot showed that P62 and LC3A/B increased in the aging group compared with the normal one. (L) Western blot showed that DRAM and GABARAP decreased in the aging group compared with the normal one. Each bar represents the mean ± SD; n = 3; *p < 0.05.