Fig. 5

Administration experiment of neutralizing antibodies of TNF-α and IL-1α with TD-GM-CSF MΦ. A Summary of the administration experiment of neutralizing antibodies; the same protocol was used with ASD-GM-CSF MΦ. B Representative images at DIV56 of immunostaining of MAP2 + dendrite co-cultured with TD-GM-CSF MΦ and simultaneous administration of neutralizing antibodies for TNF-α, IL-1α, or both, or corresponding isotype antibodies (IgG) at a concentration of 1 μg/ml. All neurons were induced from 201B7 hiPSC line, and co-cultured with macrophages of TD4. Scale bar: 100 μm. C–G Results of (C) total length of MAP2 + dendrite, D average length of MAP2 + dendrite, E max length of MAP2 + dendrite, F MAP2 + dendrite count, and G branch point count of MAP2 + dendrite. The shortening of MAP2 + dendrites observed in co-culture with TD-GM-CSF MΦ was prevented by the simultaneous administration of neutralizing antibodies. C–F One-way ANOVA test with post hoc Tukey’s multiple comparison test (C) F (3, 142) = 6.606, p = 0.0003, D F (3, 142) = 11.25, p < 0.0001, E F (3, 142) = 13.05, p < 0.0001, (F) F (3, 142) = 1.544, p = 0.2058. G Brown–Forsythe ANOVA test, F (3.00, 127.0) = 1.759, p = 0.1584, with post hoc multiple comparison test of Dunnett’s T3. n(TD-GM-CSF MΦ + IgG) = 35 fields of 15 independent dishes from five times differentiations of two control healthy hiPSC lines each, n(TD-GM-CSF MΦ + anti-IL-1α Ab) = 38 fields of 18 independent dishes from five times differentiations of two control healthy hiPSC lines each, n(TD-GM-CSF MΦ + anti-TNF-α Ab) = 37 fields of 17 independent dishes from five times differentiations of two control healthy hiPSC lines each, and n(TD-GM-CSF MΦ + 2Abs) = 36 fields of 16 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