ecis instrument applied biophysics ztheta 96 well array station  (Applied BioPhysics)

Journal: bioRxiv

Article Title: Th17 effector cytokines induce shared and distinct microglial and endothelial cell responses in post-streptococcal encephalitis

doi: 10.64898/2026.02.04.703836

Figure Lengend Snippet: a , Schematic of CD4⁺ T cell differentiation, effector cytokine production, and associated brain pathology in ROR γ t -/- mice following recurrent GAS infections. b, Heatmaps of differentially expressed genes (DEGs) related to BBB function, antigen presentation, and interferon response in olfactory bulb (OB) brain endothelial cells (BECs) from GAS-infected wild-type (WT) and ROR γ t -/- mice. Values are shown as log(z-score); significantly altered genes are indicated in black (adjusted p < 0.05). c, Gene ontology (GO) pathway enrichment analysis of transcripts upregulated and downregulated in ROR γ t -/- versus WT BECs after GAS infections. d, IFNγ concentrations in whole OB lysates from WT PBS (gray), WT GAS (blue), and ROR γ t -/- GAS (green) mice after two or five infections. Statistical comparisons were performed using one-way ANOVA (ns, p > 0.05; p < 0.05). e, Heat maps of DEGs related to antigen presentation, homeostatic/DAM programs, and cytokine/chemokine signaling in OB microglia from GAS-infected WT and ROR γ t -/- mice (log(z-score); adjusted p < 0.05 shown in black). f, GO pathway enrichment analysis of transcriptional changes in ROR γ t -/- versus WT microglia following GAS infections. g, Representative flow cytometry plots of CD74 and MHC class II (I-A/I-E) expression in WT and ROR γ t -/- microglia after GAS infections (n = 3,640 and 4,657 cells, respectively). h, Quantification of microglial surface expression of antigen presentation markers in WT PBS, WT GAS, and ROR γ t -/- GAS mice. Statistical comparisons were performed using one-way ANOVA (ns, p > 0.05; p < 0.05; * p < 0.01). i, j, Transendothelial electrical resistance (TEER) measurements in primary mouse BEC monolayers following cytokine treatment, shown as representative ECIS traces ( i ) and area-under-the-curve (AUC) quantification ( j ). Gray shading indicates treatment period. Data are mean ± SEM (n = 5 replicates from 3 independent experiments). Mixed-effects analysis ( p < 0.05; * p < 0.01). k, l, Relative transwell permeability of primary mouse BEC monolayers to albumin - AF594 following cytokine treatment. Untreated cells were used as reference, and LPS served as a positive control. Data are mean ± SEM (n = 3 independent experiments). Repeated-measures one-way ANOVA ( p < 0.05). m, n, TEER measurements in primary human brain microvascular endothelial cells (HBMECs), shown as representative ECIS traces ( m ) and AUC quantification ( n ). Data are mean ± SEM (n = 6 replicates from 3 independent experiments). Mixed-effects analysis (*** p < 0.0001).

Article Snippet: Transendothelial electrical resistance (TEER) was measured in real time using an electric cell-substrate impedance sensing (ECIS) instrument (Applied BioPhysics, ZTheta 96 Well Array Station) as previously described [ ]. mBECs and HBMECs were plated on 96-well plates containing electrode arrays (Applied BioPhysics, 96W20idf).

Techniques: Cell Differentiation, Immunopeptidomics, Olfactory, Infection, Flow Cytometry, Expressing, Permeability, Positive Control

Journal: Frontiers in Pharmacology

Article Title: Prostaglandin E 2 stimulates opposing effects on inner and outer blood-retina barrier function

doi: 10.3389/fphar.2025.1608376

Figure Lengend Snippet: PGE 2 , but not PGF 2α , strengthens hRMEC barrier function. (A) Normalized resistance ECIS results of hRMEC stimulated with 100 nM PGE 2 , 100 nM PGF 2α , or vehicle over 12 h (n = 8–16, passage 8). (B) Dose-response curve of hRMEC stimulation with 3 pM–10 μM PGE 2 or vehicle over 12 h (n = 6–12, passage 8). (C) Dose-response curve of hRMEC stimulation with 3 pM–10 μM PGF 2α or vehicle over 12 h (n = 6-8, passage 8). (D) Transwell dextran flux across hRMEC stimulated with 100 nM PGE 2 or vehicle after 24 h (n = 3, passage 8). All data represent mean ± SD shown by error bars. 1B-C were analyzed using three-parameter nonlinear regression models with the 95% confidence intervals shown. 1D was analyzed using an unpaired T-test with the P -value shown.

Article Snippet: ECIS assays were performed in an ECIS Z-Theta 96-well array station (Applied BioPhysics).

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Prostaglandin E 2 stimulates opposing effects on inner and outer blood-retina barrier function

doi: 10.3389/fphar.2025.1608376

Figure Lengend Snippet: PGE 2 -induced barrier enhancement of hRMEC is mediated by the EP4 receptor. Normalized ECIS resistance measures from hRMEC pretreated with (A) 500 nM SC-51322, (B) 500 nM PF-04418948, (C) 500 nM DG-041, (D) 500 nM L-161,982, or vehicle for 3 h followed by stimulation with 10 nM PGE 2 or vehicle over 12 h (n = 6-7, passage 8). Each antagonist is separated to an individual graph for clarity. (E) ECIS results expressed as AUC over 12 h from treatment of hRMEC with 50 nM–500 nM of EP antagonists +10 nM PGE 2 or vehicle (n = 6-7, passage 8, comparisons to 500 nM EP4 antagonist shown). (F) Dose-response curve of hRMEC pretreated with 10 pM-10 μM L-161,982 or vehicle for 3 h followed by 10 nM PGE 2 or vehicle stimulation over 12 h (n = 3-8, passage 8). (G) Transwell dextran flux across hRMEC pretreated with 1 μM L-161,982 or vehicle followed by 100 nM PGE 2 or vehicle stimulation after 24 h (n = 3, passage 8). (H) Dose-response curve of hRMEC stimulation with 1 pM–100 nM L-902,688, 20 nM PGE 2 , or vehicle over 12 h (n = 4-6, passage 8). (I) Normalized resistance ECIS results of hRMEC stimulated with 1 μM forskolin, 10 nM PGE 2 , or vehicle over 12 h (n = 5-6, passage 8). (J) Normalized resistance ECIS results of hRMEC pretreated with 1 μM KT5720, 10 μM ESI-09, 1 μM KT5720 + 10 μM ESI-09, or vehicle for 3 h followed by 10 nM PGE 2 or vehicle stimulation over 12 h (n = 5-6, passage 8). (K) ECIS results expressed as AUC over 12 h from treatment of hRMEC ± KT5720 and ESI-09 + 10 nM PGE 2 or vehicle (n = 5-6, passage 8). All data represent mean ± SD shown by error bars. 2E, 2G, and 2K were analyzed using one-way ANOVAs and Tukey’s multiple comparisons tests with P -values shown for relevant comparisons. 2F and 2H were analyzed using three-parameter nonlinear regression models with the 95% confidence intervals shown.

Article Snippet: ECIS assays were performed in an ECIS Z-Theta 96-well array station (Applied BioPhysics).

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Prostaglandin E 2 stimulates opposing effects on inner and outer blood-retina barrier function

doi: 10.3389/fphar.2025.1608376

Figure Lengend Snippet: PGE 2 , but not PGF 2α , induces RPE barrier permeability. (A) Normalized resistance ECIS results of hRPE stimulated with 100 nM PGE 2 , 100 nM PGF 2α , or vehicle over 12 h (n = 6, passage 5). (B) Dose-response curve of hRPE stimulation with 3 pM–10 μM PGE 2 or vehicle over 12 h (n = 6-9, passage 5). (C) Dose-response curve of ARPE-19 stimulation with 3 pM–10 μM PGE 2 or vehicle over 12 h (n = 5–11, passage 24). (D) Dose-response curve of ARPE-19 stimulation with 3 pM–10 μM PGF 2α or vehicle over 12 h (n = 6-9, passage 24). (E) Dose-response curve of ARPE-19 stimulation with 3 pM–10 μM PGD 2 or vehicle over 12 h (n = 4-6, passage 24). (F) Transwell dextran flux across ARPE-19 stimulated with 100 nM PGE 2 or vehicle after 24 h (n = 9, passage 24). All data represent mean ± SD shown by error bars. 4B-E were analyzed using three-parameter nonlinear regression models with the 95% confidence intervals shown. 4F was analyzed using an unpaired T-test with the P -value shown.

Article Snippet: ECIS assays were performed in an ECIS Z-Theta 96-well array station (Applied BioPhysics).

Techniques: Permeability

Journal: Frontiers in Pharmacology

Article Title: Prostaglandin E 2 stimulates opposing effects on inner and outer blood-retina barrier function

doi: 10.3389/fphar.2025.1608376

Figure Lengend Snippet: PGE 2 -induced barrier permeability of ARPE-19 is mediated in part by the EP2 receptor. Normalized resistance ECIS results of ARPE-19 pretreated with (A) 500 nM SC-51322, (B) 500 nM PF-04418948, (C) 500 nM DG-041, (D) 500 nM L-161,982, or vehicle for 3 h followed by stimulation with 100 nM PGE 2 or vehicle over 12 h (n = 4-6, passage 25). Each antagonist is separated to an individual graph for clarity. (E) ECIS results expressed as AUC over 12 h from treatment of ARPE-19 with 50 nM–500 nM of EP antagonists +100 nM PGE 2 or vehicle (n = 4-6, passage 25, comparisons to 500 nM EP2 antagonist shown). (F) Normalized resistance ECIS results of hRPE pretreated with 100 nM-1 μM PF-04418948 for 3 h followed by stimulation with 100 nM PGE 2 or vehicle over 12 h (n = 4-6, passage 5). (G) Dose-response curve of ARPE-19 pretreated with 30 pM-3 μM PF-04418948 or vehicle for 3 h followed by 100 nM PGE 2 or vehicle stimulation over 12 h (n = 3-6, passage 24). (H) Normalized resistance ECIS results of ARPE-19 pretreated with 100 nM-10 μM TG4-155 or vehicle for 3 h followed by stimulation with 100 nM PGE 2 over 12 h (n = 5-8, passage 25). (I) Dose-response curve of ARPE-19 stimulation with 1 nM–10 μM butaprost, 100 nM PGE 2 , or vehicle over 12 h (n = 6–10, passage 24). (J) Normalized resistance ECIS results of hRMEC pretreated with 300 μM IBMX or vehicle for 3 h followed by stimulation with 1 μM forskolin, 100 nM PGE 2 , or vehicle over 12 h (n = 5-6, passage 24). (K) cAMP production in ARPE-19 pretreated with 100 nM-1 μM PF-04418948 for 1 h followed by stimulation with 1 μM PGE 2 or vehicle for 15 min (n = 7-8, passage 24). (L) Normalized resistance ECIS results of ARPE-19 pretreated with 1 μM KT5720, 10 μM ESI-09, 1 μM KT5720 + 10 μM ESI-09, or vehicle for 3 h followed by 100 nM PGE 2 or vehicle stimulation over 12 h (n = 6, passage 24). (M) Magnification of KT5720 ± ESI-09 ECIS results from 6 to 12 h. (N) ECIS results expressed as AUC over 12 h from treatment of hRMEC ± KT5720 and ESI-09 + 10 nM PGE 2 or vehicle (n = 6, passage 24). All data represent mean ± SD shown by error bars. 5E, 5K, and 5N were analyzed using one-way ANOVAs and Tukey’s multiple comparisons tests with P -values shown for relevant comparisons. 5G and 5I were analyzed using three-parameter nonlinear regression models with the 95% confidence intervals shown.

Article Snippet: ECIS assays were performed in an ECIS Z-Theta 96-well array station (Applied BioPhysics).

Techniques: Permeability

Journal: Frontiers in Pharmacology

Article Title: Prostaglandin E 2 stimulates opposing effects on inner and outer blood-retina barrier function

doi: 10.3389/fphar.2025.1608376

Figure Lengend Snippet: PGE 2 does not signal off-target via a non-EP prostanoid receptor in ARPE-19. Normalized ECIS resistance measures from ARPE-19 pretreated with (A) 50–500 nM BW A868C, (B) 50–500 nM OC000459 , (C) 50–500 nM AL8810, (D) 50–500 nM CAY10441, (E) 50–500 nM daltroban, or vehicle for 3 h followed by stimulation with 100 nM PGE 2 or vehicle over 12 h (n = 4-6, passage 24). Each antagonist is separated to an individual graph for clarity. (F) ECIS results expressed as AUC over 12 h from treatment of ARPE-19 with 50 nM–500 nM of EP antagonists +100 nM PGE 2 or vehicle (n = 4-6, passage 24). (G) ECIS results expressed as AUC over 12 h from treatment of ARPE-19 with 100 nM PGE 2 , vehicle, or 50 nM–500 nM of EP antagonists alone (n = 5-6, passage 24). All data represent mean ± SD shown by error bars. 6F was analyzed using a one-way ANOVA with Dunnett’s multiple comparisons test to compare all groups to 100 nM PGE 2 treatment, significant P -values shown. 6G was analyzed using a one-way ANOVA with Dunnett’s multiple comparisons test to compare all groups to vehicle treatment, significant P -values shown.

Article Snippet: ECIS assays were performed in an ECIS Z-Theta 96-well array station (Applied BioPhysics).

Techniques: