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: Cancer discovery

Article Title: A gain-of-function p53 mutant oncogene promotes cell fate plasticity and myeloid leukemia through the pluripotency factor Foxh1

doi: 10.1158/2159-8290.CD-18-1391

Figure Lengend Snippet: (A) The ratio of sh.p53 BFP+ cells compared to sh.Ren BFP+ cells over time in leukemic lines. (B) CD11b expression in the shRNA BFP+ containing cells (n = 3) and (C) Annexin V shown by FACS 7 days after infection. Data presented as mean ± s.e.m (D) Relative expression of TP53 7 days post infection with pRS sh.TP53 in human KY821 cells. (E) CFU in human AML cells with sh.Ren or sh.TP53. Data presented as mean ± s.e.m (n = 3) (F) hCD16 expression by FACS in pRS Ctrl or pRS TP53 cells from growing in CFU assay (G) Percentage of hCD45 in the peripheral blood of mice 5 weeks post transplant in shCntr and shCntr moribund mice (black), shTP53 (blue) and shTP53 moribund mice (orange). Data are represented as mean ± s.e.m (n=5 and n=5) (H) Kaplan-Meier curve from NSG mice transplanted with equal numbers of KY821 cells transduced with wither shCntr or shTP53 (n=5 and n=5). (I) Western blot for human TP53 in cells before injection and in sorted hCD45 cells from the peripheral blood of 3 independent moribund shTP53 mice. *p <0.05, **p <0.005 ***p <0.0005. B. Kruskal-Wallis 1-way ANOVA. C, D, F, J, unpaired t-test.

Article Snippet: Cell lines The human leukemia line KY821 (JCRB cell bank) was grown in RPMI 1640 and 20% FBS (Gibco).

Techniques: Expressing, shRNA, Infection, Colony-forming Unit Assay, Transduction, Western Blot, Injection