ecis software  (Applied BioPhysics)

Journal: Science Advances

Article Title: Modulation of blood-tumor barrier transcriptional programs improves intratumoral drug delivery and potentiates chemotherapy in GBM

doi: 10.1126/sciadv.adr1481

Figure Lengend Snippet: ( A ) IF staining of CDH5 (red) and nuclei (blue) in HCMEC/D3 cells treated with BIA (1 μM, 24 hours). Scale bar, 20 μm. ( B ) TEER values of HCMEC/D3 treated with BIA upon monolayer confluence. Time point of BIA addition is indicated. ( C ) Interference RNA depletion of CDH5 in HCMEC/D3 cells and TEER values of these cells upon ECIS measurement. Western blot for CDH5 depletion confirmation is shown. β-Actin was used as a loading control. ( D ) IF images of BBB spheroids treated with indicated doses of BIA for 72 hours. Staining of F-actin (green), CDH5 (red), and nuclei (blue). Maximal projection intensity is shown from z-stack images (50 μm depth, 20 layers). Scale bar, 100 μm. ( E ) FITC-conjugated dextran (70 kDa) permeability assay in BBB spheroids. Dextran (gray) and nuclei (purple) are shown. Scale bar, 100 μm. Mean fluorescence quantification of ( F ) CDH5, ( G ) Phalloidin and ( H ) FITC-dextran from images in (D) and (E) using the ImageJ software. A.F., Alexa Fluor. Data show mean and SD, n = 4 to 5. Ordinary one-way analysis of variance (ANOVA) test. * P = 0.018, ** P = 0.0028, *** P = 0.008, and **** P < 0.0001. ( I ) Human phospho-kinase array of HCMEC/D3 cells exposed to 1 μM BIA for 24 hours. Highlighted wells related to indicated pathways. Samples were analyzed in duplicates. ( J ) Quantification of signal by ImageJ of dot blot shown in (I). Mean and SD of duplicates are shown. Two-way ANOVA analysis was performed. ** P = 0.0015, *** P = 0.005, and **** P < 0.0001.

Article Snippet: Using the ECIS Z-Theta software (Applied Biophysics), measurements were set to 4000 and 64,000 Hz every 30 min.

Techniques: Staining, Western Blot, Control, Permeability, Fluorescence, Software, Dot Blot

Journal: Frontiers in Medicine

Article Title: Serum from patients with cirrhosis undergoing liver transplantation induces permeability in human pulmonary microvascular endothelial cells ex vivo

doi: 10.3389/fmed.2024.1412891

Figure Lengend Snippet: Example plots of transendothelial resistance of human pulmonary microvascular endothelial cells (HPMECs) after stimulation with 5% human serum from healthy subjects (red) or one liver transplant patient (S1–S4, see legend). Measurements were obtained at 4,000 Hz using the ECIS ZΘ system. Resistance was normalized to the plateau value obtained just prior to stimulation at time = 0. (A) HPMECs from a 24-year-old male endothelial cell donor. (B) HPMECs from a 57-year-old female endothelial cell donor. Each plot is obtained from one experiment with all wells measured simultaneously on a 96-well plate. The error bars indicate the SD of three replicate wells. The arrows indicate the peak permeability effect.

Article Snippet: Impedance and TER were recorded using ECIS Software v.1.2.215.0 PC (Applied Biophysics, Inc.) at a sampling rate of approximately one measurement every 6 min. Once HPMECs reached confluency, as evidenced by the TER reaching a plateau (typically 24–36 h after seeding; ), half (150 μL) of the media was removed from each well and replaced with 150 μL of EGM-2 MV containing the human sera or controls.

Techniques: Permeability

Journal: Molecular Medicine Reports

Article Title: Loss of keratin 23 enhances growth inhibitory effect of melatonin in gastric cancer

doi: 10.3892/mmr.2023.13145

Figure Lengend Snippet: Dynamic monitoring and western blot analysis of KRT23-KO GC cells. (A) Dynamic monitoring by ECIS revealed the growth status of KRT23-KO AGS and SNU-216 cells and their control cells for 5 consecutive days. (B) Western blot analysis of expression and activation of proteins in KRT23-KO AGS and SNU-216 cells and their control cells. *P<0.05 and **P<0.01. KRT, keratin; KO, knockout; GC, gastric cancer; NC, negative control; p-, phosphorylated.

Article Snippet: ECIS software version 1.2.123 (Applied BioPhysics, Inc.) was already installed on the PC and connected to the ECIS instrument, ready for the time course measurement of impedance values.

Techniques: Western Blot, Expressing, Activation Assay, Knock-Out, Negative Control

Journal: Molecular Medicine Reports

Article Title: Loss of keratin 23 enhances growth inhibitory effect of melatonin in gastric cancer

doi: 10.3892/mmr.2023.13145

Figure Lengend Snippet: Cell cycle analysis and dynamic monitoring of KRT23-KO cells treated with or without MLT. (A) Flow cytometric analysis of KRT23-KO GC cells with or without 2.5 mM of MLT treatment for 24 h. Percentages of cell populations at diverse stages of the cell cycle were listed within the panels. All histograms represented mean ± standard deviation values of three biological replicates. (B) Dynamic monitoring of KRT23 knockout cells treated with or without 2.5 mM of MLT for 24 h using the ECIS method. *P<0.05, **P<0.01 and ***P<0.001 vs. the KO NC + Ctrl group; # P<0.05 and ### P<0.001 vs. the KO NC + MLT group; & P<0.05 and &&& P<0.001 vs. the KO KRT23+Ctrl group. KRT, keratin; KO, knockout; NC, negative control; MLT, melatonin; GC, gastric cancer.

Article Snippet: ECIS software version 1.2.123 (Applied BioPhysics, Inc.) was already installed on the PC and connected to the ECIS instrument, ready for the time course measurement of impedance values.

Techniques: Cell Cycle Assay, Standard Deviation, Knock-Out, Negative Control