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. 2020 Aug 27;11:1866. doi: 10.3389/fimmu.2020.01866

Figure 5.

Figure 5

mTOR signaling supports IL-1β production in macrophages. The mRNA expression of ATF4 (n = 4), Data are represented as mean ± SEM. (B) Left: Immunoblotting to detect the protein abundance of mTOR, p-mTOR, S6K1, and p-S6K1 in TGPMs (n = 3); Right: Statistically analysis the relative abundance of proteins between two groups. Data are representative of three independent experiments. (C) Representative western bolt displaying time-dependent change of protein abundance of mTOR and p-mTOR in TGPMs (n = 4). (D) The secretion of IL-1β from TGPMs in completed medium and completed medium treated with Rapamycin (10μM) (n = 3). (E) Left: Immunoblotting to detect the protein abundance of mTOR, p-mTOR, HIF-1α, p65, p-p65, Caspase1, and IL-1β; Right: Statistically analysis the relative abundance of proteins between two groups (n = 3/4). (F) The secretion of IL-1β and TNF-α from TGPMs in M1 or M1-S group treated with Rapamycin (10 μM) for 15h (n = 3). (G) The relative abundance of p-mTOR/mTOR and p-4EBP1/4EBP1 in M1 or M1-S group treated with Rapamycin (10 μM) (n = 3). (H) The secretion of IL-1β from TGPMs in M1 or M1-S group treated with MHY1485 (20 μM) for 15 h or not (n = 3). (I) The relative abundance of p-mTOR/mTOR and p-4EBP1/4EBP1 in M1-S or M1-S group treated with MHY1485 (20 μM) (n = 3). (J) The secretion of IL-1β from TGPMs in M1 or M1-S group treated with Leucine (50 μM) for 15 h or not (n = 4). M1-S: thioglycolate-elicited peritoneal macrophages (TGPMs) were stimulated with LPS (1 μg/ml) plus IFN-γ (20 ng/ml) in serine deficiency medium; M1: TGPMs were stimulated with LPS (1 μg/ml) plus IFN-γ (20 ng/ml) in completed medium. Macrophages were stimulated with LPS plus IFN-γ for 15 h except indicated. Data were analyzed with one-way ANOVA (H,J) or unpaired t-test (A,B,D–G,I) and represented as mean ± SD except indicated. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.