![Figure 3. Rescue from a disease-causing HD mutant by improving signal efficiency. (A) Brain homogenates from three individual mice from each of the three Prl-PrP(AV3) transgenic lines were compared with serial dilutions of homogenate from PrP(A117V)H mice, which served as a standard. After SDS-PAGE, the upper portion of the gel was stained with colloidal coomassie blue, whereas the lower portion was immunoblotted using 3F4 antibody. (B) Quantification of two experiments, as in A, showing the PrP(A117V)H standards (black circles) and each transgenic line (mean ± SD; n = 6). Using an expression level of 4× for PrP(A117V)H, the expression for lines 6, 10, and 11 were determined to be 2.4×, 4.7×, and 5.7×, respectively. (C) Expression levels of Prl-PrP(AV3) lines were compared with previously characterized mouse lines known to produce CtmPrP and develop neurodegeneration (Hegde et al., 1998a, 1999). The A3922 transgenic line, which expresses at 4×, served as a standard. Two amounts of each sample were loaded, and the blots were first stained for total protein with Ponceau S before immunoblotting with 13A5 antibody. The samples were run on two gels, with one set of samples (from PrP[KH-II]H) duplicated on each gel to ensure that they were directly comparable. Below the lanes are the quantified expression levels and mean age of neurodegeneration taken from either this or earlier studies. “n/a” indicates that disease is not observed in these lines. The vertical black line indicates that intervening lanes have been spliced out. (D) Kaplan-Meier survival plots for the indicated transgenic mouse lines. The normal lifespan of this strain of mouse in our facility is ∼600–800 d. The broken line indicates the age at which PrP(AV3) founders were observed to develop signs of disease (Hegde et al., 1998a). Numbers to the left of the blots in A and C indicate molecular mass in kD.](https://cdn.rupress.org/rup/content_public/journal/jcb/188/4/10.1083_jcb.200911115/6/jcb_200911115_rgb_fig3.jpeg?Expires=1773555727&Signature=cZMqByFxUptktiLlYN42cBXkZHcgUP6MW5xoBTmpcHEEp8ri4O1CLjhhTLxxgx43krU06WoMCu5hQua7Hj4cSTbxntCe9~Pue2FPtnRrdzgcsNPzQg7OJdhHnUrQeTbIzDLAn8T9iBrDfmHnvoHfAC6RsIDIVpyCdxNXxiE8wGceXOczmr6QTsw5ma1yOL1DOx39RhOJVplRDBLt10oSTBzc54KDMHiVrWk6SSEJCCp2VuI-Lgc-Enq46jTnwxQaWo3ZmxyurRKPFy5Swfh7Rdj-6y4wwHhri0Sb678EV5I1Ov1-3bVsMdOOOTBJ8ddzyjZmMg3Y~o9YNUvbt1Yblw__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Rescue from a disease-causing HD mutant by improving signal efficiency. (A) Brain homogenates from three individual mice from each of the three Prl-PrP(AV3) transgenic lines were compared with serial dilutions of homogenate from PrP(A117V)H mice, which served as a standard. After SDS-PAGE, the upper portion of the gel was stained with colloidal coomassie blue, whereas the lower portion was immunoblotted using 3F4 antibody. (B) Quantification of two experiments, as in A, showing the PrP(A117V)H standards (black circles) and each transgenic line (mean ± SD; n = 6). Using an expression level of 4× for PrP(A117V)H, the expression for lines 6, 10, and 11 were determined to be 2.4×, 4.7×, and 5.7×, respectively. (C) Expression levels of Prl-PrP(AV3) lines were compared with previously characterized mouse lines known to produce CtmPrP and develop neurodegeneration (Hegde et al., 1998a, 1999). The A3922 transgenic line, which expresses at 4×, served as a standard. Two amounts of each sample were loaded, and the blots were first stained for total protein with Ponceau S before immunoblotting with 13A5 antibody. The samples were run on two gels, with one set of samples (from PrP[KH-II]H) duplicated on each gel to ensure that they were directly comparable. Below the lanes are the quantified expression levels and mean age of neurodegeneration taken from either this or earlier studies. “n/a” indicates that disease is not observed in these lines. The vertical black line indicates that intervening lanes have been spliced out. (D) Kaplan-Meier survival plots for the indicated transgenic mouse lines. The normal lifespan of this strain of mouse in our facility is ∼600–800 d. The broken line indicates the age at which PrP(AV3) founders were observed to develop signs of disease (Hegde et al., 1998a). Numbers to the left of the blots in A and C indicate molecular mass in kD.
