Supplementary Figure 6: Characterization of conditional Rab7 knockout mice
From: Age-related myelin degradation burdens the clearance function of microglia during aging

(a) Strategy for conditional targeting of Rab7 in mice. (b) Timeline diagram for histological analysis of conditional Rab7 knockout mice. (c) Confocal image of cortex of CX3CR1+/CreERT2:Rab7flox/+: tdTomato+ mice is shown. Microglia were visualized with Iba1 in green and the number of Iba1+/tdTomato+ (red) cells were counted one week after tamoxifen injections (for 5 days with 75mg/kg body weight). tdTomato is expressed in more than 90% of the microglia. Scale bar: 300µm. (d) RT-PCR analysis of Rab7 from isolated microglia from Rab7 flox/flox (control) and Rab7flox/flox:CX3CR1+/CreERT2 (Rab7ΔMG) mice. (e) Confocal image of the cortex of control and Rab7ΔMG animals 6 weeks after tamoxifen injections is shown. Arrows point to enlarged lysosomes (Lamp1, red) within microglia (Iba1, green). Scale bar: 20µm. Quantification of number of Lamp1-positive microglia per area (n=4 mice per group, mean +/- s.d., two-way ANOVA, genotype effect: ***P<0.0001, F= 421.5, followed by Bonferroni’s post hoc test, ***P<0.001, *P=0.0425, t=3.115). Each dot represents the mean value of 3 brain slices per mouse.