Fig. 1

Human Casp6 expression in ACL and ACL/G mice. a Red safe-stained agarose gel of human CASP6 (CASP6), murine Casp6 (Casp6), or 18S RNA amplicons from hippocampus (H), cortex (Cx), cerebellum (Cb), or liver (L) of WT/WT, WT/Cre, Type I KI/WT and KI/Cre, and Type II of KI/WT and KI/Cre mice. b qRT-PCR-measured levels of murine Casp6 mRNA from WT/WT, WT/Cre, Type I KI/WT and KI/Cre, and Type II KI/WT and KI/Cre hippocampus, cortex, cerebellum, and liver mRNA normalized to 18S RNA. Data shows mean and s.e.m. Each symbol represents data from one mouse. c Western blot of hippocampal, cortical, cerebellar and liver proteins detected with LS-B477 anti-human Casp6 antibody (top panel), and Cell Signaling 9762 anti-mouse and human Casp6 (bottom panel). d Quantification of Casp6 protein levels detected by 9762 in (c) normalized to Casp6 levels in WT/WT. Data represents mean ± s.e.m. Statistical evaluations were done with one-way ANOVA (p < 0.0001) followed by Dunnett’s post-hoc analysis vs WT/WT **p < 0.01, ***p < 0.001, **** < 0.0001 e Human Casp6 immunohistological staining with LS-B477 in hippocampal CA1 (SO; stratum oriens, PCL: pyramidal cell layer, SR: stratum radiatum, SLM: stratum lacunosum molecular), cortex, corpus callosum, fimbria and columns of the fornix regions from Type I and Type II KI/WT and KI/Cre mice. Bar = 50 μm. Arrows indicate immunopositive neurons and neurites. Arrow heads indicate dot-like staining. f Red safe-stained agarose gel of PCR amplified human CASP6 transgene STOP sequence and CaMKIIα-Cre transgenes from WT/WT, WT/Cre, Type I KI/WT and KI/Cre (ACL) and Type II KI/WT and KI/Cre (ACL/G) hippocampus (H) and tail (T) DNA