VHZ is overexpressed in breast cancer. Breast cancer samples were assessed by indirect double IF) and IHC for VHZ expression. A. Double-labeling of VHZ and γ-tubulin in the same tissue section. VHZ (c) co-localized with γ-tubulin (b) at enlarged centrosomes (white arrowheads) of cells. Image c shows merged images a and b. Bar: 100 μm. B. IHC detection of VHZ or γ-tubulin in two consecutive sections of the same breast cancer sample. Positive signals showed VHZ in brown. Similar centrosomal labeling patterns of VHZ (a) and γ-tubulin localization (b) were indicated by the black arrows. Three rectangular areas boxed in overview images (a'-c', magnification ×630) were further enlarged (×5) and shown in panels a-c, respectively. Panel c showed VHZ non-expressing breast cancer sample as a negative control. C. VHZ was overexpressed throughout the cytoplasm of dispersed epithelia in some breast cancer samples. Selected sections from different breast samples were shown in overview images (a' and b'). Three rectangular areas boxed in the overview images (a'-c', × 400) were further enlarged (×5) and shown as top panels a - c, respectively. Panel c showed one of the DUSP23-negative samples. Normal breast tissue showed no signals of VHZ protein expression (S. Figure 2A). D. A full western blot with 4 pairs of human breast tissues showed that VHZ protein (16 kDa) was detected in all 4 breast tumor samples indicated, but not respective matched normal tissues (except 1 with low level). T = tumor tissue, while N = normal tissue.