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. 2010 Sep 27;207(10):2271–2281. doi: 10.1084/jem.20092401

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© 2010 Kranich et al.

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Figure 1. — Mfge8^−/− mice show accelerated disease progression. (A) B6.129-Mfge8^−/−, B6.129-Mfge8^−/+, and B6.129-Mfge8^+/+ were inoculated i.c. with 3 log LD₅₀ (n = 10 for Mfge8^−/−, n = 4 for Mfge8^+/−, and n = 11 for Mfge8^+/+) and i.p. with 3 log LD₅₀ per group of RML6 prions. The mean incubation time after i.c. inoculation was significantly accelerated in Mfge8^−/− mice (159 ± 2 d in Mfge8^−/−, 179 ± 3 d in Mfge8^+/−, and 202 ± 3.9 d in Mfge8^+/+ mice; P < 0.0001, logrank). Differences in incubation time after i.p. inoculation were less pronounced (206 ± 6 d in Mfge8^−/− and 229 ± 10 d in Mfge8^+/+ mice) but still significant (P = 0.004, logrank test). (B) Stainings of brain sections after 30 dpi (left) or at terminal stage (right) using anti-PrP antibody SAF84. Bars, 1 mm. (C) Western blot of PK-digested brain homogenates from i.c. inoculated Mfge8^+/+ and Mfge8^−/− mice using anti-PrP antibody POM1 (C, uninoculated brain homogenate). Western blot for actin on non-PK–digested sample is shown below. Densitometric quantitation of POM1 signal normalized over actin expression. Bar graph shows mean relative signal intensity (n = 4; *, P = 0.03; Student’s t test). (D) MPA of brain homogenates from Mfge8^+/+ and Mfge8^−/− mice at 120 dpi or at terminal disease (n = 6–7 mice per group; *, P = 0.034; Student’s t test) indicated in percentages of RML6 prion–inoculated brain homogenate. Mfge8^−/− mice sacrificed at 120 dpi showed a slight increase over Mfge8^+/+ mice. (E) Mfge8^+/+ and Mfge8^−/− cerebellar slices were inoculated with 3 × 3 log LD₅₀ RML6 prions or with noninfectious brain homogenate (mock), harvested after 5 wk, homogenized, and PK digested (C, brain homogenate from a terminal RML inoculated mouse used as positive control). PrP^Sc levels were analyzed by Western blotting using POM1 and normalized against actin (without PK digestion) by densitometry (bar graph; n = 4; *, P = 0.024; Student’s t test). Error bars represent SD. Histology, Western blot, and MPA results represent at least four independent experiments.