human b lymphoblastoid cell line tk6  (ATCC)

Journal: Toxicology Reports

Article Title: Genotoxicity risk assessment of a 7-hydroxymitragynine-enriched Kratom preparation: An integrated in silico and in vitro approach

doi: 10.1016/j.toxrep.2026.102206

Figure Lengend Snippet: Micronucleus test of Kratom leaf extract after 4 h exposure with S9 in TK6 cells. Results are the mean ± SD of 3 independent experiments. Statistical testing with one-way ANOVA and Tukey’s post-hoc test (* p < 0.05).

Article Snippet: The human B lymphoblastoid cell line (TK6) (CRL-8015; batch No. 70045146), purchased from ATCC, was cultured in RPMI 1640 medium supplemented with 10 % fetal bovine serum (FBS) and 1 % penicillin/streptomycin.

Techniques:

Journal: Toxicology Reports

Article Title: Genotoxicity risk assessment of a 7-hydroxymitragynine-enriched Kratom preparation: An integrated in silico and in vitro approach

doi: 10.1016/j.toxrep.2026.102206

Figure Lengend Snippet: Micronucleus test of Kratom leaf extract after 4 h exposure without S9 in TK6 cells. Results are the mean ± SD of 3 independent experiments. Statistical testing with one-way ANOVA and Tukey’s post-hoc test (* p < 0.05).

Article Snippet: The human B lymphoblastoid cell line (TK6) (CRL-8015; batch No. 70045146), purchased from ATCC, was cultured in RPMI 1640 medium supplemented with 10 % fetal bovine serum (FBS) and 1 % penicillin/streptomycin.

Techniques:

Journal: Toxicology Reports

Article Title: Genotoxicity risk assessment of a 7-hydroxymitragynine-enriched Kratom preparation: An integrated in silico and in vitro approach

doi: 10.1016/j.toxrep.2026.102206

Figure Lengend Snippet: Micronucleus test of Kratom leaf extract after 24 h exposure without S9 in TK6 cells. Results are the mean ± SD of 3 independent experiments. Statistical testing with one-way ANOVA and Tukey’s post-hoc test (* p < 0.05).

Article Snippet: The human B lymphoblastoid cell line (TK6) (CRL-8015; batch No. 70045146), purchased from ATCC, was cultured in RPMI 1640 medium supplemented with 10 % fetal bovine serum (FBS) and 1 % penicillin/streptomycin.

Techniques:

Journal: Genes & Diseases

Article Title: TGIF2-mediated HMGB3 overexpression promotes esophageal squamous cell carcinoma proliferation and metastasis through TLR3/TGF-β signaling

doi: 10.1016/j.gendis.2025.101987

Figure Lengend Snippet: HMGB3 interacts with TLR3 and triggers the Smad-dependent TGF-β signaling pathway via NF-kB signaling in esophageal squamous cell carcinoma (ESCC). (A) Immunohistochemistry analysis reveals TLR3 expression in 20 pairs of ESCC tissues and the corresponding paratumor tissues. (B) Western blotting analysis of EC9706 cells treated with different concentrations of poly (I:C) reveals that TLR3 positively regulates the Smad-dependent TGF-β pathway. (C) Quantitative PCR reveals the RNA expression correlation between HMGB3 and TLR3 in ESCC cell lines. (D) The co-immunoprecipitation test was conducted to explore the direct interaction between TLR3 and HMGB3. (E) The effect of HMGB3 down-regulation on NF-κB P65 nuclear expression in ECA109 was analyzed by Western blotting. (F) Quantitative PCR demonstrates the RNA levels of TGF-β and TLR3 after NF-κB P65 was up-regulated by plasmid or down-regulated by siRNA. (G) Western blotting analysis illustrates the protein levels of TGF-β and TLR3 after NF-κB P65 was up-regulated by plasmid or down-regulated by siRNA. (H, I) Chromatin immunoprecipitation and quantitative PCR assays demonstrate that TGF-β and TLR3 promoters could be directly bound by NF-κB P65. All data were expressed as mean ± standard deviation. Statistical significance is indicated as ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001.

Article Snippet: Afterward, before incubation with primary antibodies overnight at 4 °C, membranes were blocked with 5% non-fat milk at 37 °C for 1 h. Subsequently, membranes were treated with horseradish peroxidase-conjugated secondary antibodies against rabbit or mouse IgG (Abcam, Massachusetts, USA, 1:5000) at 37 °C for 1 h. The following primary antibodies were used to assess the expression of proteins: anti-β-actin (#3700; Cell Signaling Technology, Massachusetts, USA, 1:5000), anti-TGIF2 (#ab190152; Abcam, 1:1000), anti-p-TGIF2 and TGIF2 (#sc-390870; Santa Cruz, CA, USA), anti-HMGB3 (#ab75782; Abcam, 1:1000), anti-TLR3 (#ab62566; Abcam, 1:1000), anti-TGF-β (#ab215715; Abcam , 1:1000), anti-SMAD2/3 (#8685; Cell Signaling Technology, 1:1000), anti-SMAD2 (#5339; Cell Signaling Technology, 1:1000), anti-p-SMAD2 (#3108; Cell Signaling Technology, 1:1000), anti-SMAD3 (#9523; Cell Signaling Technology, 1:1000), anti-p-SMAD3 (#9520; Cell Signaling Technology, 1:1000), anti-extracellular signal-regulated kinase 1/2 (ERK1/2) (#4695; Cell Signaling Technology, 1:1000), anti-p-ERK1/2 (#4370; Cell Signaling Technology, 1:1000), anti-nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB) P65 (#8242S; Cell Signaling Technology, 1:1000), and anti-His-tag (#66005-1-Ig; protein-tech, 1:1000).

Techniques: Immunohistochemistry, Expressing, Western Blot, Real-time Polymerase Chain Reaction, RNA Expression, Immunoprecipitation, Plasmid Preparation, Chromatin Immunoprecipitation, Standard Deviation