Mitochondrial Dynamics in Emphysema

Human lung tissues from normal subjects and emphysema patients and mouse lung samples were homogenized in RIPA buffer with 1× Protease Inhibitor Cocktail and 1× PMSF. The lysates were centrifuged at full speed for 15 min at 4°C and analyzed by Western blotting as previously described (Lin et al., 2017 (link)). The primary antibodies used were rabbit anti-TFAM (1: 2000, Proteintech, Cat# 22586-1-AP), mouse anti-S6 ribosomal protein (1:2000, Cell Signaling Technology, Cat# 2317), rabbit anti-phospho-S6 ribosomal protein (Ser235/236) (1:2000, Cell Signaling Technology, Cat# 4856), mouse anti-OPA1 (clone 18) (1:2000, BD Transduction Laboratories, Cat# 612606), mouse anti-DLP1 (1:2000, BD Transduction Laboratories, Cat# 611113), rabbit anti-phospho-DRP1 (Ser616) (1:1000, Cell Signaling Technology, Cat# 3455S), and mouse anti-alpha-tubulin (1:3000, Developmental Studies Hybridoma Bank, Cat# 12G10).