IL-1 and TNF-α upregulate CEBPB, autoregulate their own expression, and enhance other inflammatory factors. (A) Immunoblots showing the changes in p53 and its downstream proteins when treated with idasanutlin (Ida) (1 μM), with or without or IL-1α/ IL-1β /TNF-α (10 ng/mL). (B) The immunoblot band areas from (A) were quantified using ImageJ. The area ratios (target/vinculin for cytoplasm and target/histone 3 for nuclear protein) were determined and normalized to no cytokine treatment. Significant differences were determined using multiple comparison analysis of variance tests in comparison with the no cytokine treatment group. Additional immunoblots from more patient samples are shown in supplemental Figure 5D. (C) The heat map reveals shared significantly upregulated p53 downstream genes of 3 patient samples treated with Ida alone or in combination with IL-1β (10 ng/mL), as determined by RNA-seq. Pt, patient. (D) The volcano plot depicts DE genes in 3 AML samples treated overnight with IL-1β (10 ng/mL) + Ida (1 μM) in comparison with Ida (1 μM) treatment alone. (E) Representative flow cytometry histograms showing the expression changes in the indicated protein after treatment with IL-1β or TNF-α for 2 days. CD120a, TNF-α receptor 1, also known as TNFRSF1A; CD120b, TNF-α receptor 2, also known as TNFRSF1B; CD121a, IL-1β receptor 1, as known as IL1R1. (F) Immunoblots showing the changes in CEBPB expression of primary leukemia samples from patients with different FAB subtypes when treated with 2 different concentrations of IL-1β or TNF-α (ng/mL). (G) Immunoblots showing the changes in expression of MDM2, MCL1, CASP6, BCL2, phospho–NF-κB, and/or p38 when cultured with 2 different concentrations of IL-1β or TNF-α (ng/mL) for 48 hours. (H) Area ratios (CEBPB/vinculin for cytoplasm and CEBPB/Histone 3 for nuclear) were determined and normalized to no cytokine treatment. Significant differences were determined using multiple comparison analysis of variance tests in comparison with the no cytokine treatment group.