Additional file 1 of Microglial VPS35 deficiency impairs A�� phagocytosis and A��-induced disease-associated microglia, and enhances A�� associated pathology

Additional file 1: Figure S1. Mouse breeding and experimental protocol. a Breeding schematic to obtain VPS35f/f:CX3CR1Cre���ER:5xFAD mice. b Schematic illustrating TMX injection and behavioral testing timeline. Tamoxifen (100 mg/kg) was administered (i.p.) into VPS35f/f:CX3CR1Cre���ER:5xFAD mice [as mutants (VPS35CX3CR1:5XFAD)]. In addition, tamoxifen was injected to VPS35f/f:5xFAD or vehicle (Corn oil) was injected to VPS35f/f:CX3CR1Cre���ER:5xFAD mice [as controls (Ctrl:5XFAD)]. c Double immunostaining analysis for Iba1 (green) and VPS35 (Red) immunostaining. Scale bar = 10 ��m. d Quantitative analysis of relative microglial VPS35 intensity compared with Ctrl: 5xFAD mice. (n = 3 per group, mean �� SD, **, P 0.05, Student���s t test). Figure S3. Increased reactive astrocytes in microglial VPS35 deficient 5XFAD brain. a Co-immunostaining of GFAP (green), ApoE (red) and Thio-S (gray) in the cortex of Ctrl:5XFAD and VPS35CX3CR1:5XFAD mice. b���e Quantification of GFAP+ cell density, ApoE+ cell density, average ApoE fluorescence intensity per astrocyte and relative ApoE+Thio-S+ density in the cortex of Ctrl:5XFAD and VPS35CX3CR1:5XFAD mice (n = 5 per group, mean �� SD, **, P