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Fig. 7 | BMC Medicine

Fig. 7

From: NG2 glia regulate brain innate immunity via TGF-β2/TGFBR2 axis

Fig. 7

Activation of TGF-β2/TGFBR2 signaling attenuates LPS-induced neuroinflammation. a qPCR analysis showing the reduction of IL-1β mRNA levels in primary cultured microglia treated with TGF-β2 (5, 10 ng ml−1, 4 h) in the presence or absence of LPS (50 ng ml−1). Unpaired t test (n = 3 independent experiments). b, c Representative immunoblots showing the increases in the levels of IL-1β and TNF-a protein induced by LPS were attenuated by TGF-β2 treatment in BV2 cells (n = 8 independent experiments). This process is accompanied by alterations in the levels of phospho-SMAD2 (c). n = 9 independent experiments. d qPCR analysis shows that the disruption of TGFBR2 signaling by LY2109761 exacerbates inflammatory response to LPS stimulation. BV2 cells were treated with LY2109761 (10 ng/ml) or vehicle DMSO for 2 h, followed by exposure to LPS (100 ng/ml) or saline for 6 h. Two-way ANOVA with Turkey’s test (n = 6 independent experiments). e, f Immunoblot analysis showing the increases in the levels of active-IL-1β in BV2 cells with disruption of TGFBR2 signaling by LY2109761. f Quantification of data shown in e. Two-way ANOVA with Turkey’s test. Data are expressed as mean ± SEM (n = 3–6). *P < 0.05, **P < 0.01, ***P < 0.001. g qPCR analysis of the striatum of wild-type mice that received stereotactic injection of lentivirus-TGFBR2 showed that the production of inflammatory mediators was attenuated by TGFβR2 overexpression. Unpaired t test (n = 8–9). *P < 0.05, **P < 0.01, ***P < 0.001. ***P < 0.001. h Representative western blots showing the decreases in levels of IL-1β in the striatum of mice that received stereotactic injection of lentivirus-TGFBR2. The striatum in contralateral side received injection of lentivirus-GFP served as control. Data are expressed as mean ± SEM (n = 10). Paired t test. *P < 0.05. i qPCR analysis shows the alterations in mRNA levels of IL-1β and CX3CR1 in the brain that received intracerebroventricular infusion of active form of TGF-β2. The animals received single injection of LPS (2.5 mg/kg, i.p.) 1 h after TGF-β2 infusion. Unpaired t test (n = 4–5 mice per group). **P < 0.01

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