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. 2021 Mar 12;27:e928547-1–e928547-14. doi: 10.12659/MSM.928547

Figure 2.

Figure 2

Citrus alkaline extracts (CAE) inhibited Adriamycin RD (ARD)-induced aging in A549 cells. (A, B) The analysis of senescence-associated β-galactosidase staining (SA-β-Gal). The SA-β-Gal-positive cells percentages were scored; the mean±SD of data from at least 3 separate experiments are visualized. # P<0.05 vs Blank, * P<0.05 vs ARD. Magnification: ×200. (C) Western blot analysis of PDGF-a, TNF-α, PDGF-b, MMP-7, CyclinD1, and CTGF in A549 cells. Each group was operated in triplicate, and experiments were repeated 3 times. Bar graphs show the relative quantification. # P<0.05, ## P<0.01, ### P<0.001, and #### P<0.0001 vs Blank; * P<0.05, ** P<0.01, *** P<0.001, and **** P<0.0001 vs ARD. (D) Real-time qPCR analysis of genes; relative expression of PDGF-a, TNF-α, PDGF-b, MMP-7, CyclinD1, and CTGF. # P<0.05, ## P<0.01, ### P<0.001, and #### P<0.0001 vs Blank; * P<0.05, ** P<0.01, *** P<0.001, and **** P<0.0001 vs ARD. (E) The analysis of p16 in A549 cells by western blot. Each group was assessed in triplicate, and experiments were repeated 3 times. Bar graphs show the relative quantification, ## P<0.01 vs Blank, ** P<0.01, and *** P<0.001. (F) Real-time qPCR analysis of genes and relative expression of p16. ### P<0.001 vs Blank; ** P<0.01, ***P<0.001, and **** P<0.0001 vs ARD.