Fig. 5

Detection of PSA in PCa cell lines, xenograft tumors, and human serum by AgNIS. A The mRNA expressions of PSA in PCa cell lines of LNCaP and PC-3 as retrieved from the Cancer Cell Line Encyclopedia (CCLE). B Western blots of cellular (cell lysate) and shedded (supernatant) PSA in LNCaP and PC-3, respectively, with high and low expressions. The expression and secretion of PSA in LNCaP were effectively induced and inhibited by DHT and ENZA, respectively, while no pharmacological effects were observed in PC-3. β-Actin was included as the internal loading control. C Detection of PSA in LNCaP and PC-3 cells using AgNIS upon treatment of DHT and ENZA. D Photographs of LNCaP and PC-3 human PCa xenograft mouse models and dissected tumors. Scale bars: 0.5 mm. Detection of PSA in PCa tumors from LNCaP and PC-3 xenografts by E immunoblotting and F eFIA on AgNIS. Autologous mouse prostate glands (without cross-species affinities) and PSA were included as negative and positive controls, respectively. G Serum eFIA tests of PSA in cohorts of PCa patients (n = 50) and healthy subjects (n = 19). The yellow line indicated the cut-off MFI value calculated from the ROC curve with the optimal sensitivity and specificity. H ROC curve of eFIA for PCa diagnosis. Inset: an example photograph showing the low blood/serum volume required for eFIA. I Scatter plot showing a good linear correlation between eFIA and clinical ECLIA tests (R = 0.782). J The Bland–Altman plot comparing eFIA and clinical ECLIA. 95% confidence intervals for LOA are defined by the mean ± 1.96 s.d. of differences. K Violin plots showing the correlation of serum PSA measured by eFIA with ISUP grades. Statistical analysis was performed by one-way ANOVA with Fisher's Least Significant Difference (LSD) Test