Fig. 4

Cellular senescence causes ventilation-induced diaphragm dysfunction via elevation of the p53-p21 axis. (A-D) Diaphragm protein lysates of the two groups were detected by western blot analysis, and the expression of p53 and p21 was quantified. (Before incubated with antibodies, the membranes were clipped according to target proteins molecular weight position on the instruction of the protein marker. Full-length blots are presented in Supplementary Fig. 3). (E and F) The diaphragm of the two groups was detected by qPCR to analyse the expression of p53 and p21. (G and H) Diaphragmatic tissue sections were subjected to immunofluorescence staining to analyse the expression of p21. Red represents p21; Green represents WGA; Blue represents Hoechst. Scale bar = 50 μm. (I and J) Diaphragmatic tissue sections were subjected to immunohistochemistry staining to analyse the expression of p21. Brown represents p21. Scale bar = 20 μm. Values are represented as the mean ± SD. n = 6. (*p < 0.05, **p < 0.01, ***p < 0.001)