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. Author manuscript; available in PMC: 2017 Dec 1.
Published in final edited form as: FEBS Lett. 2016 Nov 3;590(23):4381–4392. doi: 10.1002/1873-3468.12467

Figure 2. XAF1 knockdown aμgments TNF-α response.

Figure 2

(A) Transient knockdown of XAF1 in ACHN cells was achieved by infection with XAF1 shRNA lentivirus as described in Materials and Methods. XAF1-knockdown cells or cells receiving control virus were cotransfected for 24 h with an NF-κB responsive luciferase plasmid (0.4 µg) and a β-galactosidase-expression plasmid (0.4µg), followed by 20 ng/ml TNF-α treatment for another 8 h or 16 h. Luciferase activities were measured, normalized to β-galactosidase values and results presented as fold change relative to controls without TNF-α treatment. The mean and SEM were calculated based on data from three independent experiments (**p<0.01). (B) XAF1-knockdown cells or control cells were treated with 20 ng/ml TNF-α and RNA isolated at the indicated times and reverse transcribed. Quantitative real-time RT PCR was used to quantify IL-8 mRNA levels, which were normalized to GAPDH mRNA levels. Relative IL-8 mRNA levels were calculated by comparing to untreated controls. The mean and SEM were based on three independent experiments (**p<0.01; * p<0.05).