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

Fig. 4. SIRT3 deficiency abolishes PCB2-mediated protection against lung I/R injury in vivo and in vitro.

Fig. 4

a–h I/R mice were subjected to 1 h of ischemia followed by 2 h of reperfusion to induce lung I/R injury. Mice from the control group underwent sham surgical procedures. PCB2 (30 mg/kg) was injected intraperitoneally for three consecutive days before the operation. Sirt3+/+ indicates Sirt3 wild-type mice; Sirt3-/- indicates Sirt3-knockout mice. a Representative images of H&E-stained lung tissues. Scale bar = 200 μm. Black arrows indicate representative pathological damage. b Histopathological scores (Mikawa’s score) of the lung, n = 6. c The W/D ratio of the lung, n = 6. d The survival rate of the I/R mice, n = 15. e Caspase-3 activity in the lung, n = 6. f–h Bcl-2 and SIRT3 protein levels in the lung, n = 3. Sirt3-KO indicates Sirt3-knockout mice. i–m A549 cells were transfected with an siRNA targeting SIRT3 (si-SIRT3) or the negative control (si-control), as indicated, before the H/R insult. Moreover, the cells from the PCB2 groups were treated with 20 μM PCB2 for 24 h. The cells from the H/R group were subjected to 6 h of hypoxia followed by 2 h of reoxygenation. i Representative FACS dot plots of annexin V and propidium iodide dual labeling. j 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. k Representative images of TUNEL staining in A549 cells; the nuclei were stained with DAPI. Scale bar = 200 μm. l TUNEL scores. The relative number of apoptotic cells is presented as TUNEL-positive cells/DAPI (n = 6). m Caspase-3 activity in living A549 cells. n = 6. *p < 0.05. Error bars depict the standard deviations.