Figure 1
Homozygous depletion of p60 katanin leads to embryonic lethality. (A) Scheme of Katna1 gene targeting strategy. (B) Long-range PCR over 5′ targeting vector integration site. (C) Long range PCR over 3′ targeting vector integration site. (D) Quantification of mRNA levels through RT-PCR and q-PCR, n = 3 ***p < 0.001. (E,F) As expected based on the genetic approach and observed mRNA levels, quantification of katanin p60 protein expression levels through western blotting, n = 3. Data represented as mean ± S.E.M., *p < 0.05. Independent samples T-test (One-tailed). (G) Genotyping PCR, WT band 270 bp, KO-first band 236 bp. (H) Number of newborn pups per genotype. (I,J) Quantification of spastin expression levels in adult mice by western blotting, n = 3. (K) Representative images show comparable sizes of adult brains dissected from heterozygous (+/−) mice and control (+/+) littermates. (L) Nissl staining showing overall normal gross brain anatomy.  (M,N) Quantification of of PARP cleavage to analyze apoptosis, n = 3. (O) Analysis of Fluoro-jade to label degenerating neurons. DAPI labeling detects all cells. Wild-type (+/+) littermates, heterozygous (+/−), and homozygous (−/−) katanin p60 knockout mice.
