Fig 2: Immunostains for intrinsic hepatic and breast cancer markers (A) Intrinsic liver marker AFP. Serum AFP by ELISA was below the limit of detection (data not shown). (B) Liver marker Glypican-3. (C) Liver marker Hep Par 1. (D) Cancer marker HER2/Neu. (E) Cancer marker P53. (F) Cancer marker Ki-67 (nuclear).

Fig 3: Scan chart of HCC serum and normal healthy serum samples detected by the AFP/LCA protein microarray assay.HCC serum samples, 1–9; normal healthy serum sample, 10.

Fig 4: (A) Scan chart of AFP-L3 standard substances detected by the AFP/LCA-coated chips. AFP antibodies and LCA were applied to the slides. a: AFP antibody, 0.5 mg/ml; b: LCA, 4 mg/ml. Serum samples with different concentrations of AFP-L3 were used: (1–5) 100, 50, 25, 12.5, and 6.25 ng/ml; (6–9) 100, 50, 25, and 12.5 ng/ml; (10) blank control. (B) The standard curve chart and the regression equation of AFP-L3 determined using the AFP/LCA protein microarray assay.

Fig 5: (A) AFP antigen levels detected by the AFP protein microarray. AFP antibodies were applied to the slides at different concentrations: a, 1 mg/ml; b, 0.5 mg/ml; c, 0.25 mg/ml; and d, 0.125 mg/ml. Different concentrations of the AFP antigen were used: 1.80 ng/ml, 2.40 ng/ml, 3.20 ng/ml, 4.10 ng/ml, and 5.5 ng/ml. Sera from HCC patients and healthy controls were also included: 6, HCC serum; 7, HCC serum; 8, blank control; 9, healthy serum; and 10, HCC serum. (B) The standard curve chart and the regression equation for determining the AFP levels using the AFP protein microarray assay.