Fig. 5: p70S6K mediates mTOR signaling in ATF3-mediated immune regulation in vitro. | Cell Death & Disease

Fig. 5: p70S6K mediates mTOR signaling in ATF3-mediated immune regulation in vitro.

From: Loss of ATF3 exacerbates liver damage through the activation of mTOR/p70S6K/ HIF-1α signaling pathway in liver inflammatory injury

Fig. 5

Bone marrow-derived macrophages (BMMs) were isolated from WT and ATF3 KO mice and transfected with p70S6K siRNA (si-p70S6K), and then co-cultured with splenic CD4+ T cells after LPS stimulation for 6 h. Non-specific (NS) siRNA served as a control. a Western blot analysis of phosphorylated p70S6K, PHD1, HIF-1α, HMGB1, TLR4, and NF-κB in LPS-stimulated BMMs. β-actin served as an internal control. b Quantitative RT-PCR detection of TNF-α, IL-1β, IL-6, and TGF-β in LPS-stimulated BMMs. Mean ± SD (n = 3–4 samples/group). *p < 0.05, **p < 0.01. c Quantitative RT-PCR detection of Foxp3, RORγt, and IL-17A in splenic CD4+ T cells after co-culture with p70S6K siRNA or NS siRNA-pretreated BMMs. Mean ± SD (n = 3–4 samples/group). *p < 0.05, **p < 0.01. d ELISA analysis of IL-17A levels in co-culture supernatants. Mean ± SD (n = 3–4 samples/group). **p < 0.01

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