Fig. 5.

Signaling pathways altered by LRP8 knockdown in TNBC cells. RNA-Seq was conducted for SUM149 transfected with control siRNA or LRP8-targeted siRNA. a Pathway analysis and b GO analysis of the differentially expressed genes from the RNA-Seq data were conducted using DAVID. Canonical Wnt/β-catenin and MAPK signaling pathways were enriched upon LRP8 knockdown. c Western blot of active (Non-phospho) β-catenin and β-actin (loading control) in SUM149 and HCC1937 with LRP8 knockdown by siRNA. d GSEA of the RNA-Seq data using a gene set of previously reported Wnt downstream target genes. The expression of Wnt target genes was negatively correlated with the LRP8 knockdown cells. ES: enrichment score. e, f qRT-PCR for the selected Wnt target genes in SUM149 and HCC1937. Gene expression fold change between siLRP8 and siControl was calculated using the 2^−ΔΔCt method and YWHAZ was used for normalization. The results were shown as mean ± S.D. g Western blot of vinculin (loading control), phosphorylated and total protein of p38 and ERK1/2 in SUM149 and HCC1937 with LRP8 knockdown by siRNA. h Gene expression fold change of DUSPs between siLRP8 and siControl in SUM149 cells. RPKM: reads per kilobase million.