Fig. 2

Dendritic changes induced by GM-CSF MΦ. A Representative images of immunostaining of MAP2 + dendrites. All neurons were induced from 201B7 hiPSC line, and neuron only or co-cultured with macrophages of TD2 or ASD3 were used in these images. Scale bar: 100 μm. B–F Results of (B) total length of MAP2 + dendrite, C average length of MAP2 + dendrite, D max length of MAP2 + dendrite, E MAP2 + dendrite count, and F branch point count of MAP2 + dendrite. Co-culture of GM-CSF MΦ-induced shortening of dendrites compared to culturing only neurons at DIV56, and ASD-GM-CSF MΦ exerted more severe effects than TD-GM-CSF MΦ. B Brown–Forsythe ANOVA test, F (3.00, 78.20) = 107.6, p < 0.0001, with post hoc multiple comparison test of Dunnett’s T3. C Brown–Forsythe ANOVA test, F (3.00, 115.5) = 62.56, p < 0.0001, with post hoc multiple comparison test of Dunnett’s T3. D Brown–Forsythe ANOVA test, F (3.00, 109.1) = 98.52, p < 0.0001, with post hoc multiple comparison test of Dunnett’s T3. E One-way ANOVA test, F (3, 138) = 2.426, p = 0.0682, with post hoc Tukey’s multiple comparison test. F Brown–Forsythe ANOVA test, F (3.00, 93.22) = 14.30, p < 0.0001, with post hoc multiple comparison test of Dunnett’s T3. n(DIV28-neuron) = 34 fields of 16 independent dishes from five times differentiations of two control healthy hiPSC lines each, n(DIV56-neuron) = 31 fields of 14 independent dishes from five times (201B7) and 4 times (1008C15) differentiations of two control healthy hiPSC lines each, n(TD-GM-CSF MΦ) = 39 fields of 19 independent dishes from five times differentiations of two control healthy hiPSC lines each, and n(ASD-GM-CSF MΦ) = 38 fields of 18 independent dishes from five times differentiations of two control healthy hiPSC lines each. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001