Journal: bioRxiv

Article Title: 3D brain angiogenesis model to reconstitute maturation of functional human blood-brain barrier in vitro

doi: 10.1101/471334

Figure Lengend Snippet: (A) 3D confocal image of p-gp protein (red) expressed in tri-cultured vasculature at day 7. Right section view is zoomed from left 3D projection image which describes intraluminal oriented expression of p-gp (pointed as yellow arrow heads). ECs and Nucleus were labeled by anti-CD31 (green) and Hoeschst 33342 (blue), respectively. Scale bar: 20μm. (B) Conceptual description of calcein-AM efflux assay (bottom) which mimics actual effect of p-gp inhibitor on drug resistance by p-gp in vivo (top). (C) Schematic description of work flow for calcein-AM efflux assay in developed in vitro BBB with or without p-gp inhibitor Valsopodar. (D) Representative image of calcein fluorescence in each time point. Vasculature was cultured in EC mono-culture or BBB tri-culture condition with or without p-gp inhibitor pre-treatment. Scale bar: 100μm. (E) Change of calcein fluorescence intensity at the designated region on vessel wall (yellow boxes in 0hour images) every 120 minutes. (F) Quantitative comparison of change in calcein fluorescence intensity during total experiment duration (640 minutes) normalized by initial intensity. (E, F) n=65 for EC only/DMSO Control; n=70 for EC only/Valsopodar treated; n=50 for BBB Tri-culture/DMSO control; n=110 for BBB Tri-culture/Valsopodar treated. Error bars represent SEM. Statistical comparisons between analyzed values of DMSO control and Valsopodar treated group in each time point was obtained from unpaired two-tailed Student’s t-test, with the p value threshold for statistical significance set at *p<0.05; **p<0.005; ***p<0.00001.

Article Snippet: A fully perfused vascular network of EC monoculture or BBB tri-culture was pre-treated with the p-gp inhibitor Valsopodar (PSC-833; Adooq) for 10 hours at a concentration of 10μM.

Techniques: Cell Culture, Expressing, Labeling, In Vivo, In Vitro, Fluorescence, Two Tailed Test

Journal: Neuropharmacology

Article Title: Differences in amyloid-β clearance across mouse and human blood-brain barrier models: Kinetic analysis and mechanistic modeling

doi: 10.1016/j.neuropharm.2014.01.023

Figure Lengend Snippet: Total brain clearance (BCITotal(%)) of 125I-Aβ40 in control (CNTRL), with P-gp inhibitor valspodar (VALSP) or LRP1 inhibitor RAP (A), BBB clearance (BCIBBB(%)) of 125I-Aβ40 in control and inhibitor groups (B), clearance of 125I-Aβ40 by brain degradation (BCIDegradation(%)) in control and inhibitor groups (C). Data represent mean±SEM for n=4; * P<0.05, ** P<0.01 and *** P<0.001. NS is not significant.

Article Snippet: To characterize role of P-gp and LRP1, 0.5 μl of ECF containing valspodar (40 μM; XenoTech, KS), a well-established P-gp inhibitor, or RAP (1 μM; Oxford Biomedical Research, MI), an LRP1 inhibitor, were intracerebrally administered 5 min prior to 125 I-Aβ 40 injection.

Techniques:

Journal: Neuropharmacology

Article Title: Differences in amyloid-β clearance across mouse and human blood-brain barrier models: Kinetic analysis and mechanistic modeling

doi: 10.1016/j.neuropharm.2014.01.023

Figure Lengend Snippet: A) In vitro uptake of intact 125I-Aβ40 by bEnd3 and hCMEC/D3 cells. Cellular level of intact 125I-Aβ40 is expressed in fmole/mg protein following cells treatment with 0.1 nM 125I-Aβ40 without (control; CNTRL) and with RAGE (RAGE-IgG), or P-gp (valspodar; VALSP), or LRP1 (RAP) inhibitors for 15 min and 12 h in bEnd3 and hCMEC/D3, respectively. B) Percent of degraded 125I-Aβ40 in the media of bEnd3 and hCMEC/D3 cells following treatment with 0.1 nM of 125I-Aβ40, with or without inhibitor. Data represent mean±SEM from three independent experiments; *P<0.05, **P< 0.01 and *** P<0.001.

Article Snippet: To characterize role of P-gp and LRP1, 0.5 μl of ECF containing valspodar (40 μM; XenoTech, KS), a well-established P-gp inhibitor, or RAP (1 μM; Oxford Biomedical Research, MI), an LRP1 inhibitor, were intracerebrally administered 5 min prior to 125 I-Aβ 40 injection.

Techniques: In Vitro

Journal: Nature Communications

Article Title: Identification of the Notch ligand DLK1 as an immunotherapeutic target and regulator of tumor cell plasticity and chemoresistance in adrenocortical carcinoma

doi: 10.1038/s41467-025-60649-w

Figure Lengend Snippet: A Cytotoxicity of SG3199 among ADCT-701 responsive (NCI-ACC44, NCI-ACC51, and NCI-ACC56) and non-responsive (NCI-ACC40, NCI-ACC48, and NCI-ACC54) DLK1 + ACC short-term patient-derived organoids (PDOs). Cells were treated with SG3199 for 3 days ( n = 3 independent experiments for ACC44, ACC51, ACC56, and ACC48 PDOs; n = 5 independent experiments for ACC40 and ACC54 PDOs). B Drug transporter mRNA expression in DLK1 + ADCT-701 responder and non-responder PDOs as measured by quantitative RT-PCR. Results were presented as Δcycle threshold (ΔCt). Each color represents an individual PDO. R responder, NR non-responder ( n = 3 independent experiments). Unpaired t tests were used to calculate two-tailed p-values. C Flow cytometry histograms assessing ABCB1 and the number of ABCB1 molecules per cell of DLK1 + ADCT-701 responsive (NCI-ACC44, NCI-ACC51, and NCI-ACC56) and non-responsive (NCI-ACC40, NCI-ACC48, and NCI-ACC54) ACC PDOs. D ADCT-701 cytotoxicity in the NCI-ACC40 and NCI-ACC48 PDOs with and without treatment with ABCB1 inhibitors (1 μM valspodar, 10 μM elacridar and 1 μM tariquidar). Cells were treated with ADCT-701 combined with or without ABCB1 inhibitors for 7 days (data representative of n = 3 independent experiments). E Flow cytometry histograms assessing ABCB1 among 164165, 592788 and POBNCI_ACC004 PDXs (data representative of n = 2 independent experiments). F ADCT-701 cytotoxicity in 164165, 592788 and POBNCI_ACC004 PDX-derived organoids. Cells were treated with ADCT-701 for 7 days (data representative of n = 3 independent experiments). G ADCT-701 cytotoxicity in the 164165 and 592788 PDX-derived organoids treated with or without ABCB1 inhibitors (1 μM valspodar, 10 μM elacridar and 1 μM tariquidar). Cells were treated with ADCT-701 combined with or without ABCB1 inhibitors for 7 days (data representative of n = 3 independent experiments). H Volcano plot of differentially expressed genes in control tumors ( n = 4) versus post-ADCT-701 acquired resistant tumors ( n = 3) in POBNCI_ACC004 PDX. Wald test negative log 10 p-values are shown on the y -axis. I Flow cytometry histograms assessing ABCB1 among ADCT-701 resistant and control POBNCI_ACC004 PDX tumors. J ADCT-701 cytotoxicity in the ADCT-701 resistant POBNCI_ACC004 PDX-derived organoid treated with or without ABCB1 inhibitors (1 μM valspodar, 10 μM elacridar and 1 μM tariquidar). Cells were treated with ADCT-701 combined with or without ABCB1 inhibitors for 7 days (data representative of n = 3 independent experiments). Error bars represent mean values ± S.E.M. Source data are provided as a Source Data file.

Article Snippet: After 2 days of incubation, cells were treated with different concentration of ADCT-701 or SG3199 (GlpBio, #GC62691) combined with or without 1 μM valspodar (APExBIO, #A3905), 10 μM elacridar (SelleckChem, #S7772), and 1 μM tariquidar (SelleckChem, #S8028) for 7 days or 3 days respectively.

Techniques: Derivative Assay, Expressing, Quantitative RT-PCR, Two Tailed Test, Flow Cytometry, Control