Journal: Biochemistry and Biophysics Reports

Article Title: Tetracyclines downregulate the production of LPS-induced cytokines and chemokines in THP-1 cells via ERK, p38, and nuclear factor-κB signaling pathways

doi: 10.1016/j.bbrep.2015.11.003

Figure Lengend Snippet: (A) Time-dependent changes in cytokine and chemokine production in LPS-stimulated THP-1 cells. After LPS (10 μg/ml) treatment for 15, 30, 60, or 120 min without any agents and with minocycline (50 μg/ml), tigecycline (50 μg/ml), or doxycycline (50 μg/ml), cytokines and chemokines were measured using Multi Plex according to the manufacturer’s protocols. * p <0.05 compared with LPS only at 60 min. ** p <0.05 compared with LPS only at 120 min. Mino: minocycline, Tige: tigecycline, Doxy: doxycycline. (B) The rate of cytokine and chemokine production in the THP-1 cell line compared to the production of cytokines and chemokines by LPS stimulation without tetracyclines. After LPS treatment (10 μg/ml) for 30, 60, 120 or 240 min without any agents and with minocycline (50 μg/ml), tigecycline (50 μg/ml), or doxycycline (50 mg/ml), cytokines and chemokines were measured with Multi Plex.

Article Snippet: The THP-1 human monocytic leukemia cell line was purchased from RIKEN Cell Bank (Wako, Japan).

Techniques:

Journal: Biochemistry and Biophysics Reports

Article Title: Tetracyclines downregulate the production of LPS-induced cytokines and chemokines in THP-1 cells via ERK, p38, and nuclear factor-κB signaling pathways

doi: 10.1016/j.bbrep.2015.11.003

Figure Lengend Snippet: Effects of minocycline, doxycycline, and tigecycline on the modulation of NF-κB, phospho-NF-κB, IKKα/β, phospho-IKKα/β, IκBα, and phospho-IκBα in LPS-stimulated THP-1 cells. THP-1 cells were incubated without or with 10 μg/ml LPS, or with LPS plus minocycline (50 μg/ml), doxycycline (50 μg/ml), or tigecycline (50 μg/ml) for 30, 60, or 120 min. NF-κB, phospho-NF-κB, IKKα/β, phospho-IKKα/β, IκBα, and phospho-IκBα were assessed with Western blotting.

Article Snippet: The THP-1 human monocytic leukemia cell line was purchased from RIKEN Cell Bank (Wako, Japan).

Techniques: Incubation, Western Blot

Journal: Biochemistry and Biophysics Reports

Article Title: Tetracyclines downregulate the production of LPS-induced cytokines and chemokines in THP-1 cells via ERK, p38, and nuclear factor-κB signaling pathways

doi: 10.1016/j.bbrep.2015.11.003

Figure Lengend Snippet: Effects of tetracyclines (minocycline, doxycycline, and tigecycline) on the activation of phospho-ERK1/2 and phospho-p38 in LPS-stimulated THP-1 cells. THP-1 cells were incubated without or with 10 μg/ml LPS, or with LPS plus minocycline (50 μg/ml), doxycycline (50 μg/ml), or tigecycline (50 μg/ml) for 30 or 60 min. Phospho-p38 and phospho-ERK1/2 were assessed with Western blotting.

Article Snippet: The THP-1 human monocytic leukemia cell line was purchased from RIKEN Cell Bank (Wako, Japan).

Techniques: Activation Assay, Incubation, Western Blot

Journal: Biochemistry and Biophysics Reports

Article Title: Tetracyclines downregulate the production of LPS-induced cytokines and chemokines in THP-1 cells via ERK, p38, and nuclear factor-κB signaling pathways

doi: 10.1016/j.bbrep.2015.11.003

Figure Lengend Snippet: SB203580, U0126 and BAY11-7082 suppressed TNF-α and IL-8 production in LPS-stimulated THP-1 cells on treatment with or without tetracyclines. THP-1 cells were pre-incubated by SB203580 (10 μM), U0126 (5 μM) and BAY11-7082 (5 μM) for 30 min, followed treatment without or with LPS (10 μg/ml), or with LPS (10 μg/ml) plus minocycline (50 μg/ml), doxycycline (50 μg/ml), or tigecycline (50 μg/ml) for 60 min. TNF-α were measured with ELISA. * p <0.05, ** p <0.01, *** p <0.001 compared to the measurement without the inhibitor in the same group. Abbreviation; SB: SB203580, U: U0126, BAY: BAY11-7082.

Article Snippet: The THP-1 human monocytic leukemia cell line was purchased from RIKEN Cell Bank (Wako, Japan).

Techniques: Incubation, Enzyme-linked Immunosorbent Assay

Journal: Nature Communications

Article Title: E3 ubiquitin ligase RNF128 promotes Lys63-linked polyubiquitination on SRB1 in macrophages and aggravates atherosclerosis

doi: 10.1038/s41467-025-57404-6

Figure Lengend Snippet: A Subset of cells from atherosclerotic aortas was annotated into nine clusters. B Location of cell clusters with Rnf128 expression on the t-SNE plot. C Cell clusters with the expression of Rnf128 ( Rnf128 + Mφ, left), high expression of Lyz2 ( Lyz2 hi Mφ, middle), and both of these two genes ( Rnf128 + Lyz2 hi Mφ, right). Mφ, macrophages. D The expression level of Rnf128 -expressing macrophage numbers during the process of atherosclerosis. E The number of macrophages with high expression of Rnf128 and Lyz2 in atherosclerotic aortas of mice fed WD for different durations. F Colocalization analysis via immunofluorescence of RNF128 and MOMA-2 (specific for monocytes and macrophages) expression in early and advanced atherosclerotic lesions of apolipoprotein E null (ApoE −/− ) mice fed a WD for 8 weeks and 20 weeks, respectively ( n = 8 per group, hereafter n = 8). Scale bar: 100 µm. G Western blotting images of RNF128 protein levels from whole aortas and quantitative analysis ( n = 6). H Colocalization analysis of RNF128 and MOMA-2 expression in early and advanced atherosclerotic lesions from coronary atheromatous plaques of humans ( n = 8). Scale bar: 100 µm. I Immunofluorescence of RNF128 in macrophages incubated with oxidized low-density lipoprotein (oxLDL, 75 µg/mL) for different time points ( n = 6). Scale bar: 20 µm. J Western blotting (left, n = 4) and quantitative PCR (right, n = 6) of RNF128 expression in macrophages treated with oxLDL for different time points. K Western blotting (left, n = 4) and quantitative PCR (right, n = 6) of RNF128 expression in macrophages treated with a concentration gradient of oxLDL for 24 h. L , M Western blotting (left, n = 4) and quantitative PCR analysis (right, n = 6) of RNF128 expression in RAW264.7 and THP-1-derived macrophages treated with time-dependent oxLDL, respectively. The “ n ” represents the number of biologically independent samples. Data were presented as mean ± SD, Shapiro–Wilk method tested that all data were normally distributed. Unpaired two-tailed Student’s t -test was used for ( G ). One-way ANOVA followed by the Dunnett post hoc test was used for the others. Adjusted P values were provided in case of multiple-group comparisons. Source data are provided as a Source Data file.

Article Snippet: The human monocyte leukemia cell line (THP-1, KeyGene BioTech, passage No. 5 to passage No. 10) was treated with phorbol 12-myristate 13-acetate (PMA, 100 nM; HY-18739, MCE, China) for 24 h for transformation into adherent macrophages.

Techniques: Expressing, Immunofluorescence, Western Blot, Incubation, Real-time Polymerase Chain Reaction, Concentration Assay, Derivative Assay, Two Tailed Test

Journal: Infection and Immunity

Article Title: Phosphate Groups of Lipid A Are Essential for Salmonella enterica Serovar Typhimurium Virulence and Affect Innate and Adaptive Immunity

doi: 10.1128/IAI.00123-12

Figure Lengend Snippet: LPS-induced cytokine production in cultured Mono Mac 6 (MM6) and RAW264.7 cells. IL-6 (A), IL-1β (B), and TNF-α (C) in the supernatant of the MM6 cell culture stimulated with 0.1 pmol/ml LPS for 24 h were quantified by Bioplex assay. TNF-α (D) and GM-CSF (E) released into the culture supernatant of RAW264.7 cells stimulated for 24 h with 0.1 pmol/ml or 10 pmol/ml of LPS, respectively, were quantified by Bioplex assay. Means and SEM of three independent experiments, each performed in triplicate, are given. ***, P < 0.001; **, P < 0.01; *, P < 0.05 (relative to the levels obtained by stimulation with LPS from wild-type χ3761). †††, P < 0.001; ††, P < 0.01; †, P < 0.05 (relative to levels obtained by stimulation with LPS from the parent strain, χ11065 [ΔmsbB48 ΔpagL7 ΔpagP8 ΔlpxR9]).

Article Snippet: The levels of interleukin 6 (IL-6), IL-1β, and tumor necrosis factor alpha (TNF-α) in the human monocytic leukemia cell line Mono Mac 6 (MM6) (Lonza, Braunschweig, Germany) and TNF-α and granulocyte-macrophage colony-stimulating factor (GM-CSF) in the murine macrophage cell line RAW264.7 were determined to check the LPS reactogenicity ( 20 ).

Techniques: Cell Culture, BioPlex Assay