THP-1 monocyte immune response assay

Briefly, THP-1 human monocytes (TIB-202; ATCC) were cultured in RPMI-1640 (189-02025; Wako) supplemented with 10% fetal bovine serum (175012; Nichirei), penicillin (100 U/mL) (168-23191; Wako), and streptomycin (100 mg/mL) (168-23191; Wako). F. nucleatum was cultured as previously described (43 (link)). THP-1 monocytes were pretreated with 100 μM FAC (RES20400-A7; Sigma-Aldrich), 100 μM DFO (D9533; Sigma-Aldrich), or the indicated concentration of calyculin A (038-14453; Wako) for the indicated time, followed by stimulation with 100 ng/mL LPS (tlrl-eblps; InvivoGen) for the indicated time. THP-1 monocytes were differentiated into macrophages using 6 ng/mL phorbol 12-myristate 13-acetate (AG-CN2-0010; AdipoGen) for 48 hours. Differentiated cells were pretreated with 100 μM FAC, 100 μM DFO, or 5 μM TAK-242 (13871; CAYMAN) for the indicated times, followed by treatment with F. nucleatum at a multiplicity of infection of 10 for 3 hours.

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Yamane T., Kanamori Y., Sawayama H., Yano H., Nita A., Ohta Y., Hinokuma H., Maeda A., Iwai A., Matsumoto T., Shimoda M., Niimura M., Usuki S., Yasuda-Yoshihara N., Niwa M., Baba Y., Ishimoto T., Komohara Y., Sawa T., Hirayama T., Baba H, & Moroishi T. (2022). Iron accelerates Fusobacterium nucleatum–induced CCL8 expression in macrophages and is associated with colorectal cancer progression. JCI Insight, 7(21), e156802.

Publication 2022

RPMI-1640 fetal bovine serum penicillin streptomycin calyculin A tlrl-eblps TAK-242

Calyculin a Cayman Cells Fetal bovine serum Human Infection Macrophages Monocytes Penicillin Phorbol myristate acetate Streptomycin Tak 242

Corresponding Organization : Kumamoto University

Other organizations : Gifu Pharmaceutical University

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Variable analysis

independent variables

FAC (100 μM)
DFO (100 μM)
Calyculin A (indicated concentration)
LPS (100 ng/mL)
Phorbol 12-myristate 13-acetate (6 ng/mL)
TAK-242 (5 μM)
F. nucleatum (multiplicity of infection of 10)

dependent variables

Cell response to treatments

control variables

THP-1 human monocytes (TIB-202; ATCC)
RPMI-1640 medium supplemented with 10% fetal bovine serum, penicillin (100 U/mL), and streptomycin (100 mg/mL)
F. nucleatum culture conditions (as previously described)

positive control

None explicitly mentioned

negative control

None explicitly mentioned

Annotations

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