A) HP14 induces p50 degradation in the presence of TNF alpha. Briefly, cells were transfected with NFκB-TRAFTAC and after 16 h, cells were treated with increasing HP14 concentrations. After 1h, cells were treated with or without TNF-alpha for indicated times.
B) TRAFTACs induce p50 degradation within 9 h of HP14/TNF-alpha treatment.
C) Chemical structures of HP14/TNF-alpha and the inactive epimer control (HP17).
D) VHL and TRAFTAC-dependent p50 degradation. Cells were transfected with either with NFκB-TRAFTAC or control-TRAFTAC followed by the treatment of HP14/TNF-alpha or the HP17/TNF-alpha for 12 h. Cell lysates were probed for p50 and GAPDH.
E) TRAFTAC induced p50 degradation is dependent on dCas9HT7. Stable cells overexpressing dCas9HT7 and parental cells were transfected with chimeric oligo and the experiment performed as mentioned above.
F) NFκB-TRAFTAC mediated p50 degradation elicit an effect on downstream gene expression.
(Not significant (N.S); * P < 0.03; ** p<0.002; *** p<0.0002; **** p<0.0001; n: Independent biological replicates; n=2)