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. 2021 Jun 29;53(6):1092–1108. doi: 10.1038/s12276-021-00643-6

Fig. 7. The expression of NCT regulators is regulated by Sp1 expression.

Fig. 7

a Immunoblot of whole-cell lysates of HDFs at different passages while undergoing senescence. Doubling time (DTs) corresponding to passage number are shown. b HDFs were infected with a lentivirus expressing Sp1 shRNA for 14 days; cells were selected by treatment with 5 μg/ml puromycin starting 2 days postinfection. Wide-field micrographs of cells stained for SA-β-gal activity by the cytochemical (X-gal) method. Scale bar, 50 μm. c Immunoblot of whole-cell lysates of HDFs infected with a lentivirus expressing Sp1 shRNA. d ChIP analysis with an anti-Sp1 antibody using young or senescent HDFs. The binding of Sp1 to the gene promoter was assessed by qPCR. The results are representative of three independent experiments; histograms show the means ± SD (n = 3). *, P < 0.05; **, P < 0.01; and ***, P < 0.001 by Student’s t-test. e Immunoblot and ChIP analyses with an anti-Sp1 antibody using HDFs infected with a lentivirus expressing Sp1 shRNA or control shRNA. The binding of Sp1 to the gene promoter was assessed by qPCR. The details are same as those shown in (d). f Immunoblot and ChIP analyses with an anti-Sp1 antibody using HDFs transfected with Sp1 expression or empty vector. The details are same as those shown in (d). g Proposed mechanistic model of NBIS.