STATTIC (sc-202818, Santa Cruz Biotechnology, Santa Cruz, CA, USA), SP600125 (s5567) and Anisomycin (A9789) purchased from Sigma (Louis, MO, USA) were dissolved in 100% dimethyl sulfoxide to prepare a 40 mM stock, 20 mM stock and 15 mg/ml stock respectively, and stored at -20°C. Recombinant human interleukin (IL)-6 purchased from R & D Systems (206IL, Minneapolis, MN, USA) was reconstituted in sterile PBS containing 0.1% bovine serum albumen to prepare a 10 μg/ml stock and stored at -20°C. The stock solution was added in culture medium to achieve the indicated final concentrations for cell culture.
The cell proliferation was examined using a CCK-8 cell proliferation kit (Dojindo Laboratories, Kumamoto, Japan), according to the instruction from the supplier. Absorbance was measured at 450 nm with Microplate Reader (Bio-Rad, La Jolla, CA, USA). After 24 hr serum depletion (0.2% FBS), cells were subsequently incubated in 10% FBS medium for an additional 24 hr before harvest. Cell cycle assessment and data analysis were carried out referring to our previous report
[6 (link)] using FACS Calibur flow cytometry (Becton Dickinson, Franklin Lakes, NJ, USA) equipped with the ModiFit LT v2.0 software.
The cell proliferation was examined using a CCK-8 cell proliferation kit (Dojindo Laboratories, Kumamoto, Japan), according to the instruction from the supplier. Absorbance was measured at 450 nm with Microplate Reader (Bio-Rad, La Jolla, CA, USA). After 24 hr serum depletion (0.2% FBS), cells were subsequently incubated in 10% FBS medium for an additional 24 hr before harvest. Cell cycle assessment and data analysis were carried out referring to our previous report
[6 (link)] using FACS Calibur flow cytometry (Becton Dickinson, Franklin Lakes, NJ, USA) equipped with the ModiFit LT v2.0 software.
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