Figure 2.

FBXL8 and key ubiquitin components are upregulated in human primary BRCA tissues. To clarify the potential of FBXL8-associated ubiquitination, analyses were performed by both qRT-PCR (A and B) and IHC (C). In total, 32 paired primary tissues are examined by qRT-PCR (n = 64). (A) FBXL8 shows significant overexpression in carcinoma tissues, particularly in advanced stages (**, p < 0.01). (B) SAG (sensitive to apoptosis gene), a key component of SCF E3 ubiquitin ligase, was also elevated in BRCA tissues throughout all disease stages. FBXL8 or SAG gene was normalized against GAPDH housekeeping gene. Data are representative of means ± SD (n = 3). **, p < 0.01; ***, p < 0.001. (C) H&E (hematoxylin and eosin) and immunohistochemistry (IHC) staining. Inflammatory cell infiltration is indicated with yellow arrows. Protein expression levels of FBXL8 and ubiquitin are examined in both normal (n = 30) and carcinoma (n = 30) breast tissues. The corresponding quantitative results of FBXL8 and ubiquitin in IHC are shown in panels (D) and (E), respectively. Consistent with the mRNA levels, the FBXL8 proteins are significantly upregulated in breast carcinoma tissues, as the cancer advanced into the later stages. Ubiquitin is also upregulated significantly in carcinoma tissues. The corresponding clinicopathological information for qRT-PCR analysis and IHC are shown in Figure S2 and Figure S3, respectively. Brown color indicates DAB dye-stained proteins of interest. Scale bar is 100 μm, shown as the red color line (—). ***, p < 0.001.