Extended Data Fig. 7: Sclerostin accelerates Αβ production via the Lrp6/β-catenin/BACE1 pathway.
A. The Evans blue staining and analysis (n = 3 biological samples for SOST-Tg and Sost-cKO experiments, n = 4 biological samples for SOST-KI experiments). B. The western blots for the integrity markers of the blood-brain barrier (n = 3 biological samples). C. The western blots analysis for the β-catenin and BACE1 levels in cortex and hippocampus from 18-month-old male Sost-cKO and WT mice (n = 3 biological samples for each group). D. The western blots analysis for the β-catenin and BACE1 levels in cortex and hippocampus from 6-month-old male SOST-KI and WT mice (n = 3 biological samples for each group). E. The western blots analysis for the β-catenin and BACE1 levels in cortex and hippocampus from aged female Sost-KO and WT mice (n = 3 biological samples for each group). F. The western blots analysis for the β-catenin and BACE1 levels in cortex and hippocampus from 6-month-old female SOST-KI and WT mice (n = 3 biological samples for each group). G. Western blot and grey value analysis for the β-catenin after different concentrations of recombinant sclerostin treatment (n = 2 replicated experiments and n = 3 biological samples for each experiment). H-I. Western blot analysis of p-GSK-3β expression in N2a cells after r-sclerostin (200 ng/ml) treatment for 3 hours (2 replicated experiments of 2 biological replicates). J. Western blot analysis of β-catenin and BACE1 levels in mouse neuroblastoma cells (N2a cells) after treatment with r-sclerostin (200 ng/ml) for 48 hours. K. Q-PCR analysis of Bace1 mRNA expression in N2a cells at 24 hours after treatment with r-sclerostin (200 ng/ml) with or without 2 μM BIO treatment (n = 9 biological samples). Notes: Data are represented as mean ± SEM. One-way ANOVA followed by Turkey’s multiple comparisons test (G, K). Two-tailed unpaired t-test (A, I). The grey value analysis of western blots can be seen in Supplemental Fig. 5.
