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Fig. 5 | BMC Medicine

Fig. 5

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

Fig. 5

NG2 glia maintain CX3CR1 expression and attenuate LPS-induced neuroinflammation. a Administration paradigm used for lentivirus injection and DT/DTR-induced NG2 glia ablation in iDTR adult mouse brain. Lentivirus encoding CX3CR1 were stereotactically injected into the striatum of the DTRNG2 mice and counterpart control mice. The mice were maintained for 5 days after virus injection, followed by DT injection for 4 days. The mice were sacrificed after LPS (2.5 mg/kg for 4 h) challenge. b Representative graph showing relative mRNA levels of inflammatory mediators in the striatum from mice that received unilateral injection of lentivirus-empty vector or lentivirus-CX3CR1. Two-way ANOVA with Turkey’s post test. (n = 2–3 for DTRNG2 mice; n = 11 for CTRL) To analyze the effect of the CX3CR1 in the CTRL mice, two-tailed paired t test is used. c Representative western blots showing relative protein levels of IL-1ß in the striatum from mice that received unilateral injection of lentivirus-empty vector or lentivirus-CX3CR1. Paired two-tailed t test, (n = 10), **P = 0.0020. d Representative graph showing relative mRNA levels of CX3CR1 in co-cultures in which microglial and NG2 glial are mixed in the various ratios in the presence or absence of LPS (50 ng ml−1). Two-way ANOVA with Bonferroni’s post test (n = 5 per group). e Representative western blots from two separate experiments showing IL-1β expression in the co-cultures of primary NG2 glial and microglial cells at various blend ratios in the presence or absence of LPS. f Quantification of data shown in e. Two-way ANOVA with Bonferroni’s post test (n = 3 mice per group). g Transcript levels of CX3CR1 and IL-1ß evaluated by qPCR in cultured microglia which were maintained separately from NG2 glial cells in the ratio of 1:1 in the presence of NG2 glia-conditioned medium (incubated for 4 h) and LPS (50 ng ml−1). Data are expressed as mean ± SEM. Two-way ANOVA with Tukey’s post test (n = 7–10 per group). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001

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