Figure 2. Activation of the WNT signaling and downstream target genes. (A) Immunocytochemistry of WNT3 (a,d), GSK3B (b,e), and CTNNB1 (c,f) in carcinoma cells. WNT-negative HSC2 cells (a-c) and WNT-expressing TSU cells (d-f) cultured on glass slides were applied for fluorescent immunostaining. Bar = 25 µm. (B) Analysis of WNT signaling activation in carcinoma cells. Total cellular lysates or nuclear extracts from HOC313 (lane 1), TSU (lane 2), HSC2 (lane 3), or SCCKN (lane 4) were subjected to Western blot. (C) Expression of WNT target genes in carcinoma cells. Size-fractionated protein of HOC313 (lane 1), TSU (lane 2), HSC2 (lane 3), or SCCKN (lane 4) was reacted to specific antibody to c-MYC or MT1-MMP. c-MYC was localized to the nuclei of TSU cells (b), but not to HSC2 cells (a). Bar = 25 µm.