1. Validation of BACE2 substrate candidates in mouse brain.
Validation of BACE2 substrate candidates in mouse brain. (A) DNER in the soluble TBS fraction and total cell lysates of homogenized cortex from 11-mo-old WT, Bace1−/−, Bace2−/−, and dKO male mice. Actin is used as a loading control. (B) Quantification of soluble DNER in the TBS fraction showed no significant differences between the four genotypes as analyzed by one-way ANOVA (P = 1.00 for Bace1−/− versus WT, P = 1.00 for Bace2−/− versus WT, and P = 1.00 for dKO versus WT). (C) VCAM1 in the soluble TBS fraction and total cell lysates of homogenized cortex from 11-mo-old WT, Bace1−/−, Bace2−/−, and dKO male mice. Actin is used as a loading control. (D) Quantification of soluble VCAM1 in the TBS fraction showed no significant differences between the four genotypes as analyzed by one-way ANOVA (P = 0.31 for Bace1−/− versus WT, P = 0.59 for Bace2−/− versus WT, and P = 1.00 for dKO versus WT). (E, F) No significant differences were observed in full-length levels of DNER and VCAM1 in total cell lysates (one-way ANOVA of FL DNER: P = 0.19 for Bace1−/− versus WT, P = 0.25 for Bace2−/− versus WT, and P = 0.25 for dKO versus WT; one-way ANOVA of FL VCAM1: P = 0.16 for Bace1−/− versus WT, P = 1.00 for Bace2−/− versus WT, and P = 1.00 for dKO versus WT). (G) VCAM1 in the soluble TBS fraction and total cell lysates of homogenized subdissected ventral hippocampi from 4-mo-old WT, Bace2−/−, and dKO male mice. Actin is used as a loading control. (H) VCAM1 in the soluble TBS fraction and total cell lysates of homogenized subdissected subventricular zone from P16 WT, Bace1−/−, Bace2−/−, and dKO male mice. Actin is used as a loading control. VCAM1 (I) and DNER (J) in the soluble TBS fraction and membrane fractions of homogenized brain hemispheres from 4-mo-old Bace1−/− male mice treated with CpJ or vehicle. PSEN1 membrane protein was used as a loading control.
Iryna Voytyuk et al. LSA 2018;1:e
© 2018 Voytyuk et al.