Fig 1: (a) Linear response curve to increasing PSA concentrations in spiked bovine serum samples (N = 10). (b) Specificity measurements (N = 10 measurement repetitions on a single device for 10 different devices), each modified with PSA antibody and introduced to different proteins solutions: Cytochrome C, BNP, cTnT, and PSA. The results clearly show that only the specific PSA protein biomarker leads to a high fluorescent response, while the nonspecific proteins show a negligible response. (c) In vivo measurements in five human volunteers for quantification of capillary blood PSA concentration using the SiNPs device (cyan bars, N = 3), in comparison to venous blood-based ELISA measurements (blue bars). (d) Fluorescence image of the multiplex detection of two different fluorophores on the multiple sensing area device. (e) Fluorescence image of multiplex detection with three different antigens on the same chip.
Fig 2: Berberine interacts with RPS6KB1 and potentiates radiation effects. A. Logarithmically growing parental, RPS6KB1-knockout (S6K-KO), and RPS6KB1-overexpressing (S6K-OE) LNCaP cells were treated with 0, 5, 10, 15, 25, or 30 μg/ml berberine or vehicle control (methanol) for 72 h. CellTiter 96 Aqueous One solution assay was used to determine the effect on growth ability. Data presented are mean ± SD from three independent experiments. B. A ribbon representation of bilobal S6K1-berberine model highlights mode 1 of berberine (red stick) binding to ATP pocket (light blue, N-lobe; light green, C-lobe; dark blue, c-clasp; orange, P-loop; magenta; HM or hydrophobic motif; cyan sticks, interacting amino acids). C. The mode 2 of berberine (dark green stick, berberine) reveals its extensive rotation away from the canonical ATP-binding mode (mode 1) resulting in its close proximity to c-clasp region. D. An overlay of two modes of berberine-S6K1 interaction displays relative orientation of berberine in the ATP pocket and amino acid residues (cyan sticks) that stabilize the two modes. Logarithmically growing LNCaP (E) and C4–2B (F) cells at ~60–70 % confluency were pretreated with berberine (5 μg/ml) for 48 h and irradiated with 0, 0.25, or 0.5 Gy radiation. Cells were then plated (5000 cells/well) for colony formation over 18–20 days. Stained colonies with >50 cells were counted, and the surviving fraction was determined. Relative surviving fraction was calculated by normalizing to mock radiation control. Data presented are mean ± SD from two or three independent experiments with three technical replicates for each experiment. **P < 0.01, ****P < 0.0001, compared with vehicle control (two-way ANOVA). G. Two weeks after orthotopic implantation of C4–2B cells, athymic nude mice received no treatment (control), berberine alone (5 mg/kg in DMSO), radiation alone, berberine for three weeks followed by a single dose of 9 Gy radiation (CONC group), or radiation followed by berberine treatment (SEQ group; n = 6–8 per group). The experiment was terminated seven weeks after tumor cell implantation. Blood was collected to measure serum levels of PSA at the indicated time points.
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