Figure S3. 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.