RND3 Potentiates Proinflammatory Activation through NOTCH Signaling in Activated Macrophages

Figure 1

Analysis of the expression of Rnd3 in macrophages activated through TLRs and IFN-γ. (a) qRT-PCR (upper panel) and Western blot (lower panel) analysis of Rnd3 expression in peritoneal murine macrophages activated with LPS (100 ng/ml), IFN-γ (10 U/ml), or both for the indicated times. In Western blot analysis, ERK-2 expression was used as a loading reference. Means ± SD of three independent experiments are shown. One-way ANOVA/Bonferroni’s post-tests was performed. Statistical significance was determined at the level of , compared to control, untreated cells; ^#, ^### with respect to the corresponding LPS-treated cells. (b) qPCR analysis of Rnd3 expression in WT and Notch1/Notch2 knockout (dKONOTCH1/NOTCH2) peritoneal murine macrophages activated with LPS (100 ng/ml) and IFN-γ (10 U/ml) for different times. Means ± SD of three independent experiments are shown. One-way ANOVA analysis with Bonferroni’s post-tests was performed. Statistical significance was determined at the level of , with respect to corresponding untreated WT macrophages. qPCR analysis of Rnd3 expression in peritoneal murine macrophages activated with (c) LTA (2 μg/ml) or (d) Poly I:C (5 μg/ml) in the presence or not of IFN-γ (10 U/ml) at different times. Means ± SD of at least three independent experiments are shown. One-way ANOVA analysis with Bonferroni’s post-tests was performed. Statistical significance was determined at the level of , , compared to control untreated cells. (e) qPCR analysis of Rnd3 expression in human monocytes or THP1 cells activated with LPS (100 ng/ml), IFN-γ (10 U/ml), or both for the indicated times. Means ± SD of three independent experiments are shown. One-way ANOVA analysis with Bonferroni’s post-tests was performed. Statistical significance was determined at the level of , , compared to control untreated cells, ^#, ^##, ^### with respect to the corresponding LPS-treated cells. All qPCR data are referred to unstimulated macrophages set to 1. qPCR was performed in triplicate with riboprotein P0 as the internal control.

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