Fig. 10. Correlation of endogenous CA125 expression with cell-surface expression of endogenous galectin-1 in CHO and HeLa cells. CHO (A) and HeLa cells (B) were dissociated from culture plates using a protease-free buffer. Native cells were labeled with affinity-purified anti-galectin antibodies derived from a polyclonal rabbit antiserum. Cell-surface staining was analyzed by FACS using anti-rabbit secondary antibodies coupled to allophycocyanine to detect primary antibodies. Autofluorescence levels (red curves in A and B) were determined with cells treated with trypsin prior to the FACS analysis. Galectin-1 cell-surface levels are indicated in green (CHO; panel A) and blue (HeLa; panel B), respectively. Total galectin-1 expression levels in CHO and HeLa cells, respectively, were analyzed by quantifying galectin-1 in total SDS cell lysates based on a western blot analysis (C). Lanes 1 and 4 represent the material of 20,000 cells, lanes 2 and 5 represent the material of 50,000 cells and lanes 3 and 6 represent the material of 150,000 cells. The results from CHO cells are shown in lanes 1-3, the results from HeLa cells are shown in lanes 4-6. Galectin-1 was detected with an affinity-purified rabbit antiserum directed against recombinant full-length galectin-1.