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. Author manuscript; available in PMC: 2025 Oct 1.
Published in final edited form as: J Neurochem. 2023 Feb 17;168(10):3365–3385. doi: 10.1111/jnc.15770

Figure 7.

Figure 7.

C3 is required for phagocytosis of inhibitory perisomatic synapses

(a) Immunohistochemistry (IHC) for IBA1 and CD68 show decreased expression by IBA1+ cells in layer V of neocortex in C3−/− mice following Toxoplasma gondii (ME49) infection compared to C3+/+ ME49-infected mice. Magenta arrowheads point to CD68 expression with IBA1+ cells. Green arrowheads point to IBA1+ cells.

(b) Quantification of CD68 immunoreactivity percentage area coverage in cortical layer V in C3+/+ mock-, C3−/− mock, C3+/+ ME49-. and C3−/− ME49-infected brains. Each data point represents one biological replicate and bars depict mean ± standard error of the mean (SEM). Asterisks (**) indicate P < 0.01, (***) indicate P < 0.001, and (ns) P > 0.05 by one-way ANOVA with Sidak’s multiple comparison correction (n=3 mice per condition). C3+/+ mock- vs. C3−/− mock (P value = 0.5269; df = 8); C3+/+ mock- vs. C3+/+ ME49 (P value = 0.0026; df = 8); C3+/+ mock- vs. C3−/− ME49 (P value = 0.6698; df = 8). Normality assessed by F-test (F value = 20.92, P value = 0.0004, significant).

(c) Distribution plot of individual IBA1+ cells analyzed and pooled from images in (B). Bar represents mean number of CD68 signal intensity within IBA1+ cells.

(d) IHC for GAD67 and NeuroTrace in layer V of neocortex of C3+/+ mock-, C3−/− mock, C3+/+ ME49-. and C3−/− ME49-infected mice.

(e) Quantification of GAD67+ synapses on NeuroTrace+ somata in cortical layer V in C3+/+ mock-, C3−/− mock, C3+/+ ME49-. and C3−/− ME49-infected brains. Each data point represents one biological replicate and bars depict mean ± SEM. Asterisks (*) indicate P < 0.05, (**) indicate P < 0.01, and (ns) P > 0.05 by one-way ANOVA with Sidak’s multiple comparison correction (n=3 mice per condition). C3+/+ mock- vs. C3−/− mock (P value = 0.9795; df = 8); C3+/+ mock- vs. C3+/+ ME49 (P value = 0.0037; df = 8); C3+/+ mock- vs. C3−/− ME49 (P value = 0.5051; df = 8). Normality assessed by F-test (F value = 11.17, P value = 0.0031, significant).

(f) Distribution plot of individual NeuroTrace+ cells analyzed and pooled from images in (E). Bar represents mean number of GAD67+ synapses on NeuroTrace+ somata.

Scale bars in A: 40 μm and in D: 10 μm