Extended Data Fig. 4: The GATA4-NF-kB axis induces ABCA1 during senescence.
From: Lysosomal control of senescence and inflammation through cholesterol partitioning
a, Abundance of the indicated proteins analyzed by immunoblotting upon senescence induction in IMR90 cells (left, n = 3 biologically independent experiments). Abundance of the indicated mRNAs analyzed by qRT-PCR upon senescence induction in IMR90 cells (right, from 1 biologically independent experiment, two times each experiment was repeated independently with similar results). b, Abundance of the indicated proteins analyzed by immunoblotting upon senescence induction in BJ cells expressing the PmiR-146a-GFP SASP reporter (n = 1 biologically independent experiment). c, Abundance of the indicated proteins analyzed by immunoblotting upon replicative senescence in IMR90 cells (left, n = 2 and 3 biologically independent samples for PRO and SEN (REP), respectively). Abundance of ABCA1 mRNA analyzed by qRT-PCR upon replicative senescence in IMR90 cells (right, n = 2 and 3 biologically independent samples for PRO and SEN (REP), respectively, mean ± SEM, two-sided unpaired t test). PRO and SEN (REP) denote proliferating and replicative senescent IMR90 cells. d, Schematic representation of the promoter region of ABCA1 with the indicated qPCR primers (left). Potential RELA and GATA4 motifs are shown. IMR90 cells carrying a Dox-inducible (Tet-On) vector expressing HA-GATA4 were cross linked with formaldehyde and protein extracts were immunoprecipitated with either HA or RELA antibodies. DNA was eluted, the cross-links were reversed, and the DNA was analyzed by qPCR using the indicated primers. Signals obtained from ChIP were normalized to signals obtained from an input sample (right, n = 3 biologically independent samples, % Input, mean ± SEM, two-sided unpaired t test). e, Abundance of the indicated proteins analyzed by immunoblotting upon senescence induction in IMR90 cells carrying a Dox-inducible (Tet-On) vector expressing dominant negative p53 (Tet-p53DN) (left, n = 3 biologically independent samples). Abundance of the indicated mRNAs analyzed by qRT-PCR (right, n = 3 biologically independent samples, mean ± SEM, two-way ANOVA test with Tukey’s multiple comparisons test).
