Fig 1: Differences in AR subcellular localization associated with si-Ran. (A) Relative mRNA expression levels of Ran was determined using reverse transcription-quantitative polymerase chain reaction after Ran was silenced by si-Ran in the LNCaP cells. (B and C) Ran protein expression was analysed using western blotting after Ran had been silenced by si-Ran in the LNCaP cells. (D) ELISA analysis of PSA was performed in the culture medium supernatant of the different groups at 12–48 h. Subcellular localization of AR protein in the LNCaP cells treated with si-Ran in the presence or absence of (E) ad-HepaCAM and (F) ad-HepaCAM-mt was analysed using western blotting. The data are presented as the mean ± standard deviation. *P<0.05, **P<0.01 and ***P<0.001. Ad, adenovirus; AR, androgen receptor; HepaCAM, hepatocyte cell adhesion molecule; mt, mutant; NC, negative control; PSA, prostate specific antigen; si, small interference.
Fig 2: In vivo results of different measurements using the microneedle array. (a) Stabilization curves of PSA association at 10 pM (red curve) and 100 pM (blue curve) spiked-PBS solutions. The results indicate that the sensor requires approximately 60 s to achieve a differentiable signal. (b) One cycle close-up view taken once dissociation stabilization is achieved for PSA-spiked PBS buffer. (c) One cycle close-up view taken once dissociation stabilization is achieved for PSA-spiked serum. (d) Normalized response linear curves derived from (a) and (b). The dissociation phase was conducted in 5% EG in 100 µM phosphate buffer solution. Normalized reaction is in comparison to nonspiked buffer or serum, respectively. Pink data relates to nonspecific normalized response to 22 ng/mL cTnI and 21 ng/mL GFP. (e) In vivo intradermal capillary PSA concentrations were measured in four subjects using the microneedle array (green bars) compared to ELISA measurements of PSA concentration in venous blood (blue dots). (f) Multiplex experiment results of normalized response to PSA-spiked buffer from a device modified with PSA-specific antibody (aPSA, black curve) and a device modified with cTnI-specific antibody (acTnI, red curve). (g) Multiplex experiment results of normalized response to cTnI-spiked buffer from a device modified with PSA-specific antibody (aPSA, black curve) and a device modified with cTnI-specific antibody (acTnI, red curve). (i) Top: Deviation measurements performed on a single device via multiple entries to (100 pM spiked serum solution). Bottom: Variance measurements were performed between different devices via normalized response against a 100 pM spiked serum.
Fig 3: Differences in AR and Ran subcellular localization induced by ad-HepaCAM or ad-HepaCAM-mt in LNCaP cells. AR (A) and Ran (B) intracellular distribution analysis using immunofluorescence staining in the LNCaP cells with different treatments at 48 h. (C) AR and Ran protein expression levels in the cytoplasm and nucleus were detected using western blotting. (D) ELISA analysis of PSA was performed on the culture medium supernatants from different groups at 12–48 h. Magnification, ×200. The data are presented as the mean ± standard deviation. *P<0.05 and **P<0.01. Ad, adenovirus; AR, androgen receptor; H3, histone 3; HepaCAM, hepatocyte cell adhesion molecule; mt, mutant; PSA, prostate specific antigen.
Supplier Page from Abcam for Human PSA ELISA Kit (KLK3)