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. 2009 Jan 22;106(7):2200–2205. doi: 10.1073/pnas.0807611106

Fig. 6.

Fig. 6.

EerI and bortezomib down-regulate Ub-H2A to activate NOXA expression. (A) NOXA activation correlates inversely with downregulation of Ub-H2A. Whole cell extracts from JEKO-1 cells exposed to the indicated agents were subjected to immunoblotting analyses. For H2A blot, chromatin extracts from a portion of the cells were used. (B) Specificity of the anti-H2A antibody. The color panel shows a merged image (red, anti-Ub-H2A; green, anti-H2A). (C and D) Knock-down of RING2 activates NOXA expression. (C) Whole cell extracts from untransfected 293T cells or cells expressing the indicated shRNA for 48 h were analyzed by immunoblotting. The numbers indicate the normalized levels of Ub-H2A. (D) As in (C), except that HeLa cells transfected with the indicated constructs for 36 h were analyzed. (E) Schematic representation of the NOXA locus and the primers used in CHIP experiments. (F) Ub-H2A is associated with the NOXA promoter. (G) EerI and bortezomib reduce the presence of Ub-H2A in the NOXA promoter. Cells were treated for 8 h with EerI (10 μM) or bortezomib (BZM, 25 nM). (H-I) Bortezomib and EerI increase the association of Pol II with the body of the NOXA gene. (H) JEKO-1 cells exposed to bortezomib (BZM, 25 nM, 8 h) were subjected to CHIP analysis. (I) As in (H), except that cells were treated with EerI (10 μM, 8 h).