Pro-Inflammatory Cytokines and Antibodies Induce hnRNP A1 Dysfunction in Mouse Primary Cortical Neurons

Figures

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  Figure 1. IFNγ and TNFα induce A1 mislocalization and SG formation in neurons. Primary neurons were treated with IFNγ or TNFα and assessed for A1 mislocalization (magenta) and SG formation (G3BP, green). In contrast to control neurons, IFNγ and TNFα-treated neurons displayed significant A1 mislocalization and cytoplasmic SG formation. Mislocalized A1 also colocalized with SGs in both IFNγ and TNFα-treated neurons (arrows). There was no difference in A1 mislocalization or SG formation between the cytokine-treated neurons. One-way ANOVA with Tukey’s post hoc tests with *** p < 0.001. Data are plotted as the mean of each replicate +/− SEM. Scale bar 20 μm.

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  Figure 2. IFNγ and TNFα damage neurites. Primary neurons were treated with IFNγ or TNFα and examined for neurite damage. IFNγ and TNFα led to decreased neurite length as compared to control. One-way ANOVA with Tukey’s post hoc tests with *** p < 0.001. Each dot represents an individual neuron with error bars +/− SEM. Scale bar 20 μm.

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  Figure 3. Anti-A1 antibodies enter neurons, leading to A1 mislocalization and SG formation. Primary neurons were treated with anti-A1 antibodies or IgG isotype control antibodies. Anti-A1 antibodies (green) were found to enter neurons at a significantly higher rate than IgG control. Furthermore, neurons treated with anti-A1 antibodies displayed increased A1 mislocalization (magenta) and SG formation (G3BP, red). Arrows indicate colocalization of anti-A1 antibodies, mislocalized A1, and SGs. One-way ANOVA with Tukey’s post hoc tests with ** p < 0.01 *** p < 0.001. Data are plotted as the mean of each replicate +/− SEM. Scale bar 20 μm.

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  Figure 4. Anti-A1 antibodies damage neurites. Anti-A1 antibodies or IgG isotype control antibodies were added to primary neurons. IgG and anti-A1 antibodies both led to decreased neurite length; however, anti-A1 antibodies augmented neurite damage and led to significantly decreased neurite length as compared to both control and IgG-treated neurons. One-way ANOVA with Tukey’s post hoc tests with *** p < 0.001. Each dot represents an individual neuron with error bars +/− SEM. Scale bar 20 μm.