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. Author manuscript; available in PMC: 2017 Feb 9.
Published in final edited form as: Cell Metab. 2015 Dec 10;23(2):303–314. doi: 10.1016/j.cmet.2015.11.011

Figure 5. p53 Suppresses SASP Factors in MiDAS.

Figure 5

(A–C) Cells were infected with lentiviruses expressing control, SIRT3 or HSPA9 shRNAs, then a p53 shRNA (shp53) or control virus, were assayed 10 days later for HSPA9, SIRT3, p53 and p21 protein (A), SA-Bgal activity (B), and BrdU incorporation (C).

(D) RNA from WT and rho0 cells, infected with shp53 or control lentiviruses and cultured + or − pyruvate for 10 days was analyzed for IL-1A (upper panel) and IL-1B (lower panel) mRNA by qPCR.

(E) RNA from WT or rho0 cells, mock- or IR-treated, and cultured + or − pyruvate for 10 days was analyzed as in (D).

(F) NF-κB reporter activity in cells cultured as in (E).

(G) ChIP assays for RelA and CBP binding at the IL1A promoter in WT and rho0 cells cultured + or − pyruvate for 7 days.

(H) WT and rho0 cells were cultured + or − pyruvate and infected with control or shRelA-expressing lentiviruses, and mRNA levels were measured by qPCR.

(I) WT and rho0 cells cultured + or − pyruvate were analyzed for IL-10 and TNF-α mRNAs by qPCR. Bar graphs indicate mean + SEM.