parental cell line  (InvivoGen)

Journal: Microbiology Spectrum

Article Title: BT1549 coordinates the in vitro IL-10 inducing activity of Bacteroides thetaiotaomicron

doi: 10.1128/spectrum.01669-24

Figure Lengend Snippet: B. theta VPI-5482 conditioned media stimulates human and murine TLR2. ( A ) Mouse TLR2 reporter HEK293 cells or the parental cell line (null2) were stimulated with B. theta VPI-5482 conditioned media or TYG control media at the indicated doses for 20 h and TLR2 stimulation was assessed by quantification of secretion of embryonic alkaline phosphatase (SEAP)-mediated hydrolysis of detection media which can be measured by absorbance at 620 nm. Background signal was corrected by subtracting null2 absorbance values from mTLR2 absorbance values for each individual treatment group. ( B–D ) Human TLR2 ( B ), TLR2/6 ( C ), and TLR2/1 ( D ) reporter HEK293 cells, or the parental cell line, null1 for hTLR2, and TLR2KO-TLR1/6 for hTLR2/6 and hTLR2/1 cells, were stimulated with B. theta VPI-5482 conditioned media or TYG control media at the indicated doses for 20 h and TLR2 stimulation was assessed by quantification of secretion of embryonic alkaline phosphatase (SEAP)-mediated hydrolysis of detection media which can be measured by absorbance at 620 nm. Background signal was corrected for by subtracting null1 absorbance values from hTLR2 absorbance values, or hTLR2KO-TLR1/6 absorbance values from either hTLR2/1 or hTLR2/6 absorbance values for each individual treatment group. Data points represent the mean and the error bars represent the standard deviation, n = 2 technical replicates per dose ( A–D ). Graph is representative of three experiments ( A–D ). Statistical significance was determined using a Student’s t test on the area under the curve ( A–D ). **** P ≤ 0.0001.

Article Snippet: Parental cell lines null1 (Invivogen, Cat #hkb-null1), null2 (Invivogen, Cat #hkb-null2), and the control cell line for hTLR2/1 and hTLR2/6 reporter lines, human TLR2KO-TLR1/6 (Invivogen, Cat #hkb-htlr2k16; cells are deficient in TLR1 and TLR6) were also used.

Techniques: Control, Standard Deviation

Journal: Phytomedicine : international journal of phytotherapy and phytopharmacology

Article Title: Cytotoxicity of dioncophylline A and related naphthylisoquinolines in leukemia cells, mediated by NF-κB inhibition, angiogenesis suppression, G2/M cell cycle arrest, and autophagy induction.

doi: 10.1016/j.phymed.2023.155267

Figure Lengend Snippet: Fig. 1. Cytotoxic effect of naphthylisoquinolines, Cpds. 1–4, by the resazurin assay. (A) Dose-response curves of dioncophylline A and its derivatives, with the positive control doxorubicin on CCRF-CEM and CEM-ADR5000 cells. (B) Cytoxicity of dioncophylline A on normal healthy cells (PBMC) and HEK-Blue™ Null1 cells upon dioncophylline A treatment. The data are the result of mean values ± SD of three independent experiments.

Article Snippet: The NF-κB SEAP reporter parental cell line HEK-BlueTM Null1 was purchased from InvivoGen (San Diego, CA, USA) and cultured under the recommended conditions.

Techniques: Resazurin Assay, Positive Control

Journal: Phytomedicine : international journal of phytotherapy and phytopharmacology

Article Title: Cytotoxicity of dioncophylline A and related naphthylisoquinolines in leukemia cells, mediated by NF-κB inhibition, angiogenesis suppression, G2/M cell cycle arrest, and autophagy induction.

doi: 10.1016/j.phymed.2023.155267

Figure Lengend Snippet: Fig. 9. Immunofluorescence visualizes the NF-κB translocation in HEK-Blue™ Null1 cells. TNF-α enhanced NF-κB translocation from the cytoplasm to the nucleus in the absence of dioncophylline A. However, TNF-α induced NK-κB translocation was inhibited by pre-treatment with dioncophylline A (1, 5, and 10 µM) for 24 h. An AF7000 widefield fluorescence microscope at 40 × magnification (scale bars = 10 µm) was used for visualization.

Article Snippet: The NF-κB SEAP reporter parental cell line HEK-BlueTM Null1 was purchased from InvivoGen (San Diego, CA, USA) and cultured under the recommended conditions.

Techniques: Immunofluorescence, Translocation Assay, Fluorescence, Microscopy