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. 2022 Jul 11;13(7):594. doi: 10.1038/s41419-022-05051-w

Fig. 5. PCB2-mediated pulmonary protection depends on the enzymatic activity of SIRT3.

Fig. 5

A549 cells were pretreated with 3-TYP (50 μM, 12 h) and/or PCB2 (20 μM, 24 h) in the corresponding groups before H/R treatment. Cells in the pcDNA3.1, SIRT3 and SIRT3-H248Y groups were transfected with empty plasmid, SIRT3 expression plasmid or SIRT3-H248Y plasmid, respectively, and then subjected to H/R injury. All groups of cells were subjected to 6 h of hypoxia followed by 2 h of reoxygenation to induce H/R injury. a–c SIRT3 protein levels, n = 3. d, e Cell viability, n = 6. f and h Representative images of TUNEL staining in A549 cells; the nuclei were stained with DAPI. Scale bar = 200 μm. g and i TUNEL scores. The relative number of apoptotic cells is presented as TUNEL-positive cells/DAPI (n = 6). j and m Caspase-3 activity in living A549 cells. n = 6. k and n Representative FACS dot plots of annexin V and propidium iodide dual labeling. l and o FACS analysis of the apoptotic rate in A549 cells. The sum of the annexin V-positive population and propidium iodide-positive population is shown. n = 3. *p < 0.05, n.s indicates no significance. Error bars depict the standard deviations.