Fig 1: BACE2 but not BACE1 cleaves SEZ6 and SEZ6L in a pancreatic β-cell line. a sSEZ6 and sSEZ6L were detected in the supernatant and full-length SEZ6 and SEZ6L in the lysate of the pancreatic β-cell line MIN6 upon BACE1 and BACE2 knock-down by siRNA (siB1, siB2). As a control, cells were treated with non-silencing control siRNA (siCon). Densitometric quantitations of the Western blots are shown, (*; p < 0.05, two-tailed Mann-Whitney test n = 4). b BACE1 (green lines) and BACE2 (blue lines) activity were quantified in enzymatic (solid lines) and cellular (dotted lines) models after pharmacological inhibition with nonselective (inhibiting both BACE1 and BACE2, compounds B and C) and BACE1-selective inhibitors (compound A). Soluble Aβ42 as well as sSEZ6 and sSEZ6L were detected in the supernatant of the neuroblastoma cell line SK-N-BE(2) or in the MIN6 respectively, as indicated. Data were standardized to low and high controls within each assay. Data represent biological duplicates with two or more technical replicates
Fig 2: Proteomic analysis of CSF from BACE DKO and WT mice. a Volcano plot of proteomic analysis of BACE1 and BACE2 double knockout (BACE DKO) and WT mouse CSF. The minus log10 transformed t-test p-values are plotted against the log2 transformed label-free quantification intensity ratios of BACE DKO and WT CSF for every relatively quantified protein. Proteins with a t-test p-value < 0.05 are shown as red circles. Already known BACE substrate candidates with a p-value < 0.05 are marked with gray filling. Proteins that remain significant after Benjamini-Hochberg false discovery rate correction (FDR < 0.05) have bold letters (SEZ6 and SEZ6L). b Detection of sSEZ6 and sSEZ6L in mouse CSF. Densitometric quantitation of the Western blot is shown, (**; p < 0.01, one-way ANOVA followed by two-tailed Student’s t-Test, n = 3). The dotted line indicates that the samples were loaded onto the same blot, but not next to each other
Supplier Page from Enzo Life Sciences, Inc. for BACE2 (soluble) (human), (recombinant) (His-tag)