Fig 1: (a) Investigation of different antigen incubation time on GPC-1 biosensor. (b) Modified self-assembled monolayer system (3-MPA & DTT) based GPC-1 biosensor (Biosensor 1) on DPV measurements of multiple concentrations of GPC-1 antigen in undiluted human serum. (c) SAMSA modified antibody based GPC-1 biosensor (Biosensor 2) on DPV measurements of multiple concentrations of GPC-1 antigen in undiluted human serum. (d) The calibration curve based on Biosensor 1 and Biosensor 2. (e) Exhibition of change of current response at different concentrations of GPC-1 antigens based on two different biosensors. (f) Interference study based on HE4 antigen.
Fig 2: HE4 suppresses Erk1/2 phosphorylation in CD8+ and CD56+ cells via DUSP6 induction. Two-color flow cytometric analysis of PBMC following 24-hr incubation with PBS (CTR), rHE4 (0.01 mg /mL) and BCI (1 mM) as indicated. (A) 2-D scatterplots of phosphor-Erk1/2 (Alexa Fluor 647) and CD8 or CD56 (FITC) are shown. (B) Mean ± SEM from analyses of phosphor-Erk1/2 positive cells from four individual donors are shown in the bar graph. (C) Immunoblotting for phosphor-Erk1/2 in CD56+ NK92MI, CD8+ TALL-104 and CD4+ H9 cells following 1-h incubation with the conditioned media from 24-h PBMC culture with rHE4 (0.01 mg/mL) and BCI (1 mM) in the indicated combinations. Blots of total Erk1/2 are shown as loading controls. (D) Bar graph represents the relative band densities to controls. Mean ± SEM are shown (n = 4). (E) DUSP6 concentrations in cell lysates and culture supernatants (SN) after 24 h incubation with or without rHE4 (0.01 mg/mL). Mean ± SEM are shown (n = 4). *p < 0.05, **p < 0.01, n.s., not significant.
Fig 3: HE4 upregulates expression of DUSP6 in PBMCs. (A) DUSP6 transcription in response to a 6-hr incubation with 0.01 mg/mL rHE4 (HE4) or equivalent amount of PBS (CTR) were evaluated by triplicated trials of real time PCR using PBMCs from four individual donors. (B) Two-color flow cytometric analysis of PBMC following 24-h incubation with 0.01 mg/mL of rHE4 (HE4) or vehicle (CTR). 2D-scatterplots of DUSP6 (Alexa Fluor 647) and CD3 or CD56 (FITC) are shown. Numbers on the plots indicate mean percentages of the cell populations of the each tetrameric area from four independent experiments. (C) Bar graphs from flow cytometric analyses using PBMCs from four individual donors. The mean ± SEM are shown. *p < 0.05, **p < 0.01.
Fig 4: HE4 upregulates expression of DUSP6 in peripheral CD8+ T cells. Two-color flow cytometric analysis of PBMCs following 24-h incubation with 0.01 mg/mL of rHE4 (HE4) or vehicle (CTR). (A) 2D-scatterplots of DUSP6 (Alexa Fluor 647) and CD4 or CD8 (FITC) are shown. Numbers on the plots indicate mean percentages of the cell populations of the each tetrameric area from four independent experiments. (B) A bar graph from flow cytometric analyses using PBMCs from four individual donors. The mean using ± SEM are shown. *p < 0.01.
Supplier Page from Abcam for Recombinant Human HE4 protein