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2 protocols using c25 140 hy 120934

1

Signaling Pathway Activation Assay

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LPS (E. coli, serotype 055:B5), anti-Flag antibody (F3165), anti-Myc antibody (M4439) and anti-β-actin antibody (A2228) were purchased from Sigma. Poly(I:C), R-848 and ODN 1585 were from Enzo Life Sciences. DAPI (D1306) were purchased from Invitrogen. Antibody to p-IKKβ (2694), IKKβ (2678), p-JNK (4668), JNK (9252), p-ERK (4376), ERK (4695), p-IκBα (9246), IκBα (4812), MyD88 (4283), HA (3724), TRAF6 (4743; immunofluorescence analysis), and Sufu (2522) were purchased from Cell Signaling. Anti-His (sc-8036), anti-TRAF6 (sc-8409) (immunoprecipitation and immunoblot analysis), anti-IRAK1 (sc-5288) and anti-IRAK4 (sc-374349) were purchased from Santa Cruz. Goat anti-mouse IgG (31430) and goat anti-rabbit IgG (31460) were purchased from Thermo Fisher Scientific. C25-140 (HY-120934) was purchased from MedChemExpress.
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2

Cell Culture and Chemical Compounds

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HEK293T, MCF-7, and MDA-MB-231 cells were obtained from American Type Culture Collection. All cell lines were maintained in Dulbecco’s Modified Eagle’s Medium (DMEM) containing 10% fetal bovine serum (FBS), 100 units/mL penicillin, and 100 μg/mL streptomycin. GSK3326595 (HY-101563) and C25-140 (HY-120934) were purchased from MedChemExpress (Monmouth Junction, NJ, USA).
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