A Conserved Role for Human Nup98 in Altering Chromatin Structure and Promoting Epigenetic Transcriptional Memory (original) (raw)
Figure 2
Human HLA-DRA exhibits epigenetic transcriptional memory.
(A, Top) Schematic of activation and reactivation time courses. For activation, HeLa cells were mock treated for 72 h and then treated with IFN-γ. For reactivation, cells were first treated with IFN-γ for 24 h, washed and split into fresh medium, cultured for 48 h without IFN-γ, and then treated again with IFN-γ. (Bottom) RT qPCR on RNA harvested from cells at the indicated times during activation and reactivation. The levels of HLA-DRA mRNA were quantified relative to _β_-ACTIN. The positions of all qPCR products for human genes are shown in Figure S3. Error bars represent the standard error of the mean from five experiments. (B, Top) Schematic of treatment regime. (Bottom) Cells were fixed and harvested at the indicated times and ChIP was performed using anti-RNAPII (8WG16). The recovery of promoter and coding sequences for HLA-DRA, CIITA, and GAPDH was quantified by qPCR relative to input. (C) Cells were treated as in panel B, fixed, and ChIP was performed using anti-phospho-Ser5 CTD (4h8). The recovery of promoter and coding sequences for HLA-DRA, CIITA, and GAPDH was quantified by qPCR relative to input. For panels B and C, error bars represent standard error of the mean from three experiments.