Figure S7.

Formation of a functional microglial network, related to Figure 6

(A) Representative recording demonstrating the formation of a microglial network upon the intracranial injection of α-syn fibrils in Cx3cr1^GFP animals in vivo.

(B) Quantification of the number of connected cells per individual cells and the percentage of microglia being integrated into a cellular network upon intracranial injection of α-syn fibrils in Cx3cr1^GFP animals in vivo. n = 2 animals per group with three to four randomly chosen areas that were analyzed for network formation. Interconnected microglia were counted manually. A total of at least 42 microglia were analyzed.

(C) Quantification of process movement velocity of microglia recorded by 2-photon imaging with and without intracranial injection of α-syn fibrils in Cx3cr1^GFP animals (left panel). Quantification of process movement velocity of microglia not transmitting or transmitting α-syn aggregated to neighboring cells upon intracranial injection of α-syn fibrils. n = 25-30 individual processes were quantified.

(D) Representative recording and 3D reconstruction of the cells in Figure 6F demonstrating the formation of a microglial network (Cx3cr1^GFP, green) upon the injection of α-syn (red).

(E) Representative recording and 3D reconstruction of distant microglia (Cx3cr1^GFP, green) containing α-syn (red).

(F) Representative time-lapse recording of microglia (Cx3cr1^GFP, green) demonstrating that α-syn (red) is shuffled back into the cell soma when cells could not share the burden of α-syn by attaching to neighbor cells.

(G) Quantification of particles that underwent transfer from one cell to another for their traveling distance and particle transfer velocity for cells shown in Figure 6F and Figure S14D.

(H) Quantification of particles which transfer to a neighboring cell was unsuccessful for their traveling distance and particle transfer velocity for cells shown in Figure S14E and S14F.

All graphs are presented as mean ± SEM and were analyzed by t test. ^∗∗∗∗p < 0.0001, ^∗∗p < 0.01.

Scale bars: 20 μm.