CD45.1 mice were engrafted with heterozygous
CX3CR1/GFP+/− bone marrow as described
previously (Lu et al., 2011 (link); Marker et al., 2013 (link)). Briefly, 8-week-old
CD45.1 host mice were lethally irradiated with 9 Gy of gamma radiation from a
cesium source. Mice were head shielded to prevent any radiation-induced
neuroinflammation or damage (Ajami, Bennett,
Krieger, Tetzlaff, & Rossi, 2007 ; Mildner et al., 2007 (link)). 8- to 12-week-old
CX3CR1/GFP+/− donor mice were euthanized by
CO2 asphyxiation and the bone marrow from the femurs and tibias
were collected in Dulbecco’s PBS (dPBS; Cellgro; 21–030-CV). Red
blood cells were lysed with ACK lysing buffer (Lonza; 10–548E), and the
remaining bone marrow cells were washed twice in sterile dPBS and resuspended at
a concentration of 5 × 107 cells/ml. 200μl of this cell
suspension was then injected into the tail veins of the irradiated host mice.
The mice recovered for 4 weeks prior to Tat injection.
CX3CR1/GFP+/− bone marrow as described
previously (Lu et al., 2011 (link); Marker et al., 2013 (link)). Briefly, 8-week-old
CD45.1 host mice were lethally irradiated with 9 Gy of gamma radiation from a
cesium source. Mice were head shielded to prevent any radiation-induced
neuroinflammation or damage (
Krieger, Tetzlaff, & Rossi, 2007
CX3CR1/GFP+/− donor mice were euthanized by
CO2 asphyxiation and the bone marrow from the femurs and tibias
were collected in Dulbecco’s PBS (dPBS; Cellgro; 21–030-CV). Red
blood cells were lysed with ACK lysing buffer (Lonza; 10–548E), and the
remaining bone marrow cells were washed twice in sterile dPBS and resuspended at
a concentration of 5 × 107 cells/ml. 200μl of this cell
suspension was then injected into the tail veins of the irradiated host mice.
The mice recovered for 4 weeks prior to Tat injection.
