Fig. 1: RFX7 mediates DDIT4 up-regulation by p53 and stress.
From: p53-mediated AKT and mTOR inhibition requires RFX7 and DDIT4 and depends on nutrient abundance
A Genome browser snapshot of the DDIT4 gene locus. Upper black tracks display publicly available p53 binding signals from Nutlin-3a-treated U2OS [40] and HCT116 [39] cells. Gray and orange tracks display RFX7 binding signals from respective dimethyl sulfoxide (DMSO) and Nutlin-3a-treated U2OS, HCT116, and RPE-1 cells [31]. B ChIP-qPCR of p53 and RFX7 binding to GAPDH (negative control), MDM2 (p53 positive control), and DDIT4 from U2OS cells treated with 10 µM Nutlin-3a or DMSO solvent control. Mean and standard deviation is displayed. Statistical significance obtained through a two-sided unpaired t-test, n = 3 technical replicates. C RT-qPCR data of DDIT4 and RFX7 in 10 µM Nutlin-3a and DMSO control-treated U2OS cells transfected with two different control siRNAs (siCtrl) and two different siRNAs against RFX7. Normalized to siControl#1 DMSO and ACTR10 control gene. Mean and standard deviation is displayed. Statistical significance obtained through a two-sided unpaired t-test, n = 9 replicates (three biological with three technical each). D Western blot analysis of RFX7, DDIT4, p53, and actin (loading control) levels in U2OS, HCT116, and RPE-1 cells transfected with siControl, siRFX7, or siTP53 and treated with DMSO solvent control, 10 µM Nutlin-3a (N3A), 5 nM Actinomycin D (AD), and 1 µM Doxorubicin (Dox).
