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Figure 4 | Journal of Neuroinflammation

Figure 4

From: TGFβ signalling plays an important role in IL4-induced alternative activation of microglia

Figure 4

TGFβ1 up regulates the IL4Rα. Primary microglia were treated with TGFβ1 (1 ng/ml) for different time points and the cells were either harvested for analysing IL4Rα mRNA and protein levels, or fixed with 4% paraformaldehyde (PFA) for IL4Rα immunostaining. Quantitative RT-PCR showed that treatment with TGFβ1 increased IL4Rα mRNA levels starting after 1 hour and peaking at around 2 hours. Afterwards mRNA levels decreased again and returned to basal levels at 24 hours (A). Western blotting showed IL4Rα protein expression starting to increase after treatment with TGFβ1 for 1 hour and peaking at around 6 to 12 hours after treatment (B). After treatment with TGFβ1 for 6 and 12 hours, IL4Rα immunoreactivity (red) was increased and cell morphology changed towards a ramified phenotype compared to control cells. After 24 hours, the IL4Rα immunoreactivity (red) was decreased but the cells still presented a ramified shape with long processes (C). Scale represents 20 μm. Treatment of primary microglia with TGFβ1 (n = 5) had no effect on IL4 mRNA levels (D). Analysis of TGFβ1-mediated changes in microglial cytokine release (n = 2) demonstrated no differences in IL4 levels after TGFβ1 treatment (E). Data (A,B, D) are presented as means ± standard error: **P < 0.01, ***P < 0.001 (one-way analysis of variance).

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