Figure 3. Expression of SNAP-23 protein is reduced by half in Snap23 ^fl/wt and Snap23 ^Δ/wt mice.

(A) Snap23 ^fl/wt heterozygous mice were mated and twelve two-week old pups from this mating were genotyped and analyzed for SNAP-23 protein expression. Genotyping of tail DNA was performed using PCR Primer set 2 from tail DNA. A single small PCR fragment (266 bp) is present in Snap23 ^wt/wt pups, and double fragments (266 and 400 bp) is present in Snap23 ^fl/wt pups. No homozygous Snap23 ^fl/fl pups were obtained when more than 50 pups were analyzed from Snap23 ^fl/wt heterozygous matings. For immunoblot analysis, whole brain was solubilized in modified RIPA lysis buffer and protein levels were analyzed by immunoblotting (WB) as indicated antibodies. (B) Snap23 ^Δ/wt heterozygous mice were mated and pups from this mating were genotyped and their brains were analyzed for expression of SNAP-23 and other SNARE proteins. Genotyping of tail DNA was performed using PCR Primer set 2. A single small PCR fragment (266 bp) is present in Snap23 ^wt/wt pups, and double fragments (266 and 492 bp) is present in Snap23 ^Δ/wt pups. No homozygous Snap23 ^Δ/Δ pups were ever obtained from Snap23 ^Δ/wt heterozygous matings. Whole brains were solubilized and analyzed by immunoblotting (WB) using the indicated antibodies.